Novel pharmaceutical compositions comprising an antibody which binds the human anti-mullerian hormone receptor type ii

ABSTRACT

The invention relates to novel pharmaceutical compositions including, as active ingredient, an antibody binding the human anti-Müllerian hormone type II receptor (AMHR-II) and an anticancer agent, as well as the therapeutic applications of these compositions.

The present invention relates to novel pharmaceutical compositionscomprising, as active ingredient, an antibody binding the humananti-Müllerian hormone type II receptor (AMHR-II), as well as thetherapeutic uses of these compositions.

The human anti-Müllerian hormone is a glycoprotein of 560 amino acids, amember of the TGF-β family. It is a hormone secreted by the Sertolicells of the fetal testis, which causes degeneration of the Müllerianduct. It is expressed in the adult in the Sertoli cells and Leydig cells(testis) and the granulosa cells (ovary). It plays a role in theactivity of the adult ovary in the regulation of folliculogenesis.

The anti-Müllerian hormone type II receptor (AMHR-II) is a peptide of573 amino acids and possesses serine-threonine kinase activity. It isinvolved in the regression of the Müllerian duct associated with thedevelopment of the human reproductive system. The Müllerian ductatrophies in men, where it only forms the prostatic vesicle and thesessile hydatid, but persists in women, where it gives rise to theFallopian tubes, uterus and the greater part of the vagina. Thisreceptor is frequently expressed on human ovarian epithelial tumourcells.

International application WO 2008/053330 describes a murine 12G4monoclonal antibody directed against AMHR-II for treating ovariancancers.

International application WO 2011/141653 describes mutated humanized12G4 antibodies, or fragments thereof, possessing an affinity at leastequal to that of the corresponding unmutated chimeric antibody,specificity with respect to the AMHR-II receptor, and not eliciting animmune reaction.

The purpose of the present invention is to propose a therapeuticalternative that is advantageous for patients with a pathology connectedwith the human anti-Müllerian hormone type II receptor (AMHR-II).

A pathology associated with the human anti-Müllerian hormone type IIreceptor (AMHR-II) may in particular be: ovarian cancer, in particularmetastatic ovarian cancer, serous cancer, hypernephroma, endometrioid,colloidal epithelium, it may also be: prostate cancer, germ cell cancer,endometrial cancer, mixed Müllerian malignant tumour of the uterus,leiomyosarcoma, and endometrial stromal sarcoma.

Ovarian cancer is the main cause of gynaecological cancers and is thefifth commonest cause of mortality from cancer in woman. It has thefollowing three histological origins:

-   -   the surface epithelium (epithelial tumour with various        subtypes), which represents 85-90% of ovarian cancers,    -   sexual cords/stroma (granulosa tumour (3% of total ovarian        cancers)), which represent about 10% of ovarian tumours,    -   germ cells, which represent 5% of ovarian cancers.

It is generally asymptomatic during the initial stages, hence itsnickname “silent killer” (La Marca A., Volpe A. The Anti-Mullerianhormone and ovarian cancer. Human Reproduction Update, Vol. 13, No. 3pp. 265-273, 2007).

There are four stages and prognoses (FIGO classification: InternationalFederation of Gynaecology and Obstetrics) for which the survival ratedecreases considerably from stage 2:

Stage I: Tumour limited to the ovaries (5-year survival: 90-70%),

Stage II: Tumour in one or two ovaries and extends to pelvis (5-yearsurvival: 70-40%),

Stage III: Tumour in one or two ovaries, extending outside the pelvis(5-year survival: 20%),

Stage IV: Distant metastases excluding peritoneal metastases (5-yearsurvival: <10%),

(Fauci, Braunwald et al. Principles of internal medicine. Harrison's17th edition/National Cancer Institute cancer.gov/CNGOF (French NationalColleges of Gynaecologists and Obstetricians).

Regarding ovarian cancer, the main strategies used for treatment aresurgery and chemotherapy, in particular as first-line treatment, such asa mixture of carboplatin and paclitaxel.

Monoclonal antibodies have also recently been developed such ascetuximab, which is directed against the epidermal growth factorreceptor (EGFR, Ozols R. F. et al., Focus on epithelial ovarian cancer,Cancer Cell. 2004, January; 5(1): 19-24). Other monoclonal antibodiesare currently in phase III, such as abagovomab directed against CA-125,avastin directed against vascular endothelial growth factor (VEGF-A), orfarletuzumab directed against folate receptor alpha (FRA).

The purpose of the present invention is to propose a therapy directedagainst a target different from the antibody targets currentlydeveloped. The invention offers the advantage of proposing a treatmentagainst various pathologies associated with AMHR-II. Moreover, in thecase of ovarian cancer, the invention offers the possibility of atherapy that is more effective than the reference therapy for reducingtumour volume, thus allowing a quicker improvement in the patient'scondition.

This purpose is achieved by means of a composition according to theinvention.

The present invention relates to a pharmaceutical compositioncomprising, as active ingredient, in combination with a pharmaceuticallyacceptable vehicle,

an anticancer agent, and

an antibody binding the human anti-Müllerian hormone type II receptor(AMHR-II).

In the invention, the term “antibody” refers to an immunoglobulin, amultimeric protein consisting of 4 chains, i.e. 2 light chains and 2heavy chains, each comprising a variable region and a constant region.More precisely, each light chain consists of a variable region (V_(L))and a constant region (C_(L)). Each heavy chain consists of a variableregion (V_(H)) and a constant region consisting of three constantdomains C_(H1), C_(H2) and C_(H3). The domains C_(H2) and C_(H3) make upthe domain Fc. The variable region of the light chain consists of threeregions determining recognition of the antigen (complementaritydetermining region, CDR) surrounded by four framework domains. Thethree-dimensional folding of the variable region is such that the 3 CDRsare exposed on the same side of the protein and allow formation of aspecific structure recognizing a defined antigen.

An “anticancer agent” is defined as any molecule that can eitherinterfere with the biosynthesis of macromolecules (DNA, RNA, proteins,etc.) or inhibit cellular proliferation, or lead to cell death byapoptosis or cytotoxicity for example. Among the anticancer agents,there may be mentioned alkylating agents, topoisomerase inhibitors andintercalating agents, anti-metabolites, cleaving agents, agentsinterfering with tubulin, monoclonal antibodies.

A “pharmaceutically acceptable vehicle” refers to a non-toxic materialthat is compatible with a biological system such as a cell, a cellculture, a tissue or an organism.

According to a particular aspect, the invention relates to apharmaceutical composition comprising, as active ingredient, incombination with a pharmaceutically acceptable vehicle, an anticanceragent and an antibody binding AMHR-II, in which said antibody bindingAMHR-II is a polyclonal antibody.

The term “polyclonal antibody” denotes a mixture of antibodies, capableof recognizing various antigenic determinants of a target protein.

According to another particular aspect, the invention relates to apharmaceutical composition comprising, as active ingredient, incombination with a pharmaceutically acceptable vehicle, an anticanceragent, and an antibody binding AMHR-II, in which said antibody is amonoclonal antibody, and preferably a chimeric or humanized 12G4monoclonal antibody.

A “monoclonal antibody” is defined as an antibody that only recognizes asingle antigenic determinant of the target protein.

By “chimeric monoclonal antibody” is meant an antibody in which thevariable regions of the light chains and heavy chains belong to aspecies different from that of the constant regions of the light chainsand heavy chains. By extension or usage a chimeric antibody refers to anantibody with constant portions of human origin.

The chimeric antibodies according to the invention may be prepared usingthe techniques of genetic recombination. For example, a chimericantibody may be produced by constructing a chimeric gene comprising anucleotide sequence of complementary DNA (cDNA) or a genomic sequencewith introns coding for the variable region of the heavy chain of amurine monoclonal antibody, joined to a nucleotide sequence coding forthe constant region of the heavy chain of a human antibody, and byconstructing a chimeric gene comprising a nucleotide sequence coding forthe variable region of the light chain of a murine monoclonal antibody,joined to a nucleotide sequence coding for the constant region of thelight chain of a human antibody. By transfecting said chimeric genes, byprotoplast fusion or any other technique, into a cell line, of murinemyeloma for example, production of mouse-human chimeric antibodies bythe transformed cells is obtained. It is the document Morrison et al.,Proc. Natl. Acad. Sci. U.S.A., 81, pp. 6851-55 (1984) that described thepreparation of such antibodies for the first time. The documentsBoulianne, G. L. et al., Nature, 312: 643-646 (1984), Sun, L. K., etal., Proc. Natl. Acad. Sci. USA (1984), 214-218, U.S. Pat. No.4,816,567, U.S. Pat. No. 6,331,415, U.S. Pat. No. 6,808,901 and EP125023 could also be used as reference by a person skilled in the art,as well as Bobrzecka, K., et al., Immunology Letters 2, pp 151-155,which describes a procedure of splitting of the interchain disulphidebridges of the immunoglobulins followed by ordered rearrangement ofthese same disulphide bridges in order to obtain antibodies formed fromrabbit Fab fragments and human Fc fragments.

Another possible approach for the preparation of chimeric antibodies, asdescribed in document FR 2 641 468, is to graft Fab′ fragments of amurine monoclonal antibody onto human polyclonal immunoglobulins, inparticular IgG, or onto Fc fragments, using a coupling agent, forexample a diimide. Chimeric antibodies of the type Ig-Fab′ (alsodesignated Fab′-Ig), Fc-Fab′ or (Fab′)2 may thus be obtained. Suchchimeric antibodies are characterized by grafting the whole of the Fab′fragment, and not only the variable portions.

Alternatively, other authors have described the production of monovalentchimeric antibodies by grafting Fab′ fragments of polyclonal antibodiesonto IgG or onto Fc fragments (G. T. Stevenson et al., Med. Oncol. &Tumor, 1985, Pharmacother., Vol. 1, No. 4, 275-278, 1984).

Homologous recombination in vivo between the portions of the genescoding for the constant regions of the light chains and heavy chains ofa murine immunoglobulin by portions of the genes coding for the constantregions of the light chains and heavy chains of a human immunoglobulinis also a means that may be used in order to obtain such antibodies(U.S. Pat. No. 5,204,244 or U.S. Pat. No. 5,202,238). This is not anexhaustive list.

By “humanized monoclonal antibody” is meant an antibody in which some orall of the sequences of the regions involved in antigen recognition (thehypervariable regions (CDR: Complementarity Determining Region), andsometimes certain amino acids of the FR regions (Framework regions)),are of non-human origin (preferably murine) whereas the sequences of theconstant regions and variable regions not involved in antigenrecognition are of human origin.

The humanized antibodies according to the invention may be prepared bywell-known techniques, such as that described for the first time in thedocument by Jones et al., Nature, 1986, 321-522-525. This involvedreplacing the hypervariable regions (CDRs) of a human antibody withhypervariable regions of murine origin, both at the level of the lightchains but also of the heavy chains. This technique, now well known to aperson skilled in the art under the name “CDR grafting”, is described innumerous documents such as Singer et al., J. Immun. 150: 2844-2857(1993), Riechman et al., Nature, Vol. 332, 323: 326 (1988), or in U.S.Pat. No. 5,225,539; U.S. Pat. No. 5,585,089; EP 0682040, which may alsobe used as reference.

However, most of the humanized antibodies produced by grafting of theCDR regions have reduced affinity relative to a murine antibody, owingto the major role of certain amino acids of the framework regions in thespatial positioning of the non-human amino acids including the CDRs aswell as in the binding to the antigen. That is why today a personskilled in the art quite often replaces, in the human receiving Ig, notonly the CDRs, but also the residues of the framework regions that maycontribute to the binding site of the antigen.

Another technique for humanizing antibodies is the technique of graftingthe specific determining residues (Specificity Determining Region, SDR),which consists of no longer grafting the whole of the CDR regions, butonly the SDR regions of the non-human antibody in the human variableregions (Tamura et al., J Immunol. 2000; 164: 1432-41). The SDR regionsare defined as the regions of the CDRs in direct contact with theantigen (Padlan et al. (1995), FASEB J. 9: 133-139). This techniquetherefore requires identification of the SDRs. This may be done, forexample, by determining the 3D structure of the antigen-antibodycomplex, using the database of the SDRs already identified(http://paradox.harvard.edu/sdr), or else by comparing the humanvariable sequences with those of the non-human species, using computersoftware such as DomainGapAlign, CLUSTALW2, CLUSTALX, BLAST or FASTA.

Another alternative for obtaining humanized antibodies consists ofgrafting regions called “abbreviated CDRs” (“grafting of abbreviatedCDRs”). It involves grafting the SDR regions and some adjacent residues,upstream and downstream of the sequence. The documents by De Pascalis etal., The Journal of Immunology, 2002, 169: 3076-3084; Kashmiri Syed V. Set al., Methods, Volume 36, Issue, May 2005, Pages 25-34 will be able tobe used as reference.

The composite humanization technology developed by Antitope (WO2006082406) is a CDR grafting technique that considers the frameworkregions independently and aims to select the human equivalentsseparately in such a way that the presentation of the residuesinteracting with the antigen are better conserved in their orientation.

The so-called resurfacing technique, “variable domain resurfacing”, alsocalled “veneering”, as developed by ImmunoGen (U.S. Pat. No. 5,639,641)or Xoma (EP 0571613, U.S. Pat. No. 5,766,886, U.S. Pat. No. 5,770,196,U.S. Pat. No. 5,821,123, U.S. Pat. No. 5,869,619) may also be used. Thistechnology consists of giving a human “profile” to a mouse variabledomain by replacing the residues exposed on the surface in the frameworkregions of the murine antibodies with the residues usually found on thesurface of the human antibodies. The documents by Roguska et al., ProcNatl Acad Sci USA 1994; Mark G. E. et al. (1994) in Handbook ofExperimental Pharmacology Vol. 113: The pharmacology of monoclonalAntibodies, Springer-Verlag, pp 105-134 may also serve as reference.

The Germliner™ platform developed by AvantGen may also be used(http://www.avantgen.com/AvantGensTechnologiesandServices.pdf). Thismakes it possible to obtain humanized antibodies in which only CDR3s areof non-human origin.

This is not an exhaustive list. Obtaining said humanized antibodieswill, moreover, preferably be coupled to an affinity maturation process.

The production of the humanized 12G4 monoclonal antibody is described indetail in international application WO 2011/141653. The antibodiesdescribed in the invention are isolated and purified. These antibodiesare mature, i.e. they possess an ad hoc three-dimensional structureallowing them to recognize the antigen, and possess all thepost-translational modifications essential to their antigenicrecognition, in particular glycosylation and the formation ofintramolecular and intermolecular disulphide bridges.

According to another aspect, the invention relates to a fragment of amutated humanized 12G4 monoclonal antibody as defined above, selectedfrom the group of fragments consisting of: Fc, Fab′-SH, Fd, Fv, Fab,F(ab′)2, Fab′, dsFv, scFv, Sc(Fv)2, diabody, triabody or tetrabody oralso nanobody.

According to an even more particular aspect, the invention relates to apharmaceutical composition comprising, as active ingredient, incombination with a pharmaceutically acceptable vehicle, an anticanceragent, and an antibody binding AMHR-II, in which the chimeric orhumanized 12G4 monoclonal antibody is mutated, and comprises at leastone mutation in the light and/or heavy chain.

According to an even more particular aspect, the invention relates to apharmaceutical composition comprising, as active ingredient, incombination with a pharmaceutically acceptable vehicle, an anticanceragent, and an antibody binding AMHR-II, in which the chimeric orhumanized 12G4 monoclonal antibody is mutated, comprises at least onemutation in the light and/or heavy chain, and has an affinity forAMHR-II characterized by a K_(D) preferably less than 10⁻⁷ M, inparticular less than 10⁻⁸M, in particular in the range from 10⁻⁹M to10⁻¹¹ M.

More particularly, the present invention relates to a pharmaceuticalcomposition comprising, as active ingredient, in combination with apharmaceutically acceptable vehicle, an anticancer agent, and amonoclonal antibody binding AMHR-II, in which said monoclonal antibodyis a humanized 12G4 antibody, or a fragment of humanized 12G4 monoclonalantibody, said humanized 12G4 monoclonal antibody being mutated, andcomprises at least one mutation in the light and/or heavy chain, saidmutated antibody possessing an affinity for the human anti-Müllerianhormone type II receptor (AMHRII) characterized by a K_(D) preferablyless than 10⁻⁷ M, in particular less than 10⁻⁸ M, in particular in therange from 10⁻⁹ M to 10⁻¹¹ M.

By “mutated humanized 12G4 monoclonal antibody” is meant a humanized12G4 monoclonal antibody in which at least one mutation was carried outin the variable region of the light chain and/or the constant region ofthe light chain and/or the variable region of the heavy chain or theconstant region of the heavy chain. A mutated humanized 12G4 antibody,in a composition according to the invention, has an affinity at leastequal to that of the corresponding unmutated chimeric antibody, aspecificity with respect to AMHR-II and does not elicit an immunereaction.

Even more particularly, the present invention relates to apharmaceutical composition comprising, as active ingredient, incombination with a pharmaceutically acceptable vehicle, an anticanceragent, and a monoclonal antibody binding AMHR-II, in which said mutatedhumanized monoclonal antibody comprises or is constituted by:

a) a light chain comprising or constituted by:

-   -   a variable region the amino acid sequence of which is        represented by SEQ ID NO: 1 or SEQ ID NO: 2, and    -   a constant region the amino acid sequence of which is        represented by SEQ ID NO: 3 or by a sequence having at least 80%        homology with SEQ ID NO: 3        b) a heavy chain comprising or constituted by:    -   a variable region the amino acid sequence of which is        represented by SEQ ID NO: 4, or SEQ ID NO: 5, and    -   a constant region the amino acid sequence of which is        represented by SEQ ID NO: 6 or by a sequence having at least 80%        homology with SEQ ID NO: 6,        in which the humanized 12G4 monoclonal antibody comprises at        least one mutation in the light and/or heavy chain, and has a        K_(D) for the human anti-Müllerian hormone type II receptor        (AMHR-II) preferably less than 10⁻⁷ M, in particular less than        10⁻⁸ M, in particular in the range from 10⁻⁹ M to 10⁻¹¹ M.        According to an even more particular aspect, the invention        relates to a pharmaceutical composition comprising, as active        ingredient, in combination with a pharmaceutically acceptable        vehicle, an anticancer agent, and an antibody binding AMHR-II,        in which the chimeric or humanized 12G4 monoclonal antibody is        mutated, and comprises at least one mutation in the light and/or        heavy chain. The references of the amino acid sequences of the        different portions of the antibodies are presented in the        following table:

Sequence references Sequence reference in in the application presentinvention WO 2011/141653 Humanized 12G4 antibody, SEQ ID NO: 1 SEQ IDNO: 2 light chain, variable region, without leader Humanized 12G4antibody, SEQ ID NO: 2 SEQ ID NO: 4 light chain, variable region, withleader Humanized 12G4 antibody, SEQ ID NO: 3 SEQ ID NO: 6 light chain,constant region, Humanized 12G4 antibody, SEQ ID NO: 4 SEQ ID NO: 8heavy chain, variable region, without leader Humanized 12G4 antibody,SEQ ID NO: 5 SEQ ID NO: 10 heavy chain, variable region, with leaderHumanized 12G4 antibody, SEQ ID NO: 6 SEQ ID NO: 12 heavy chain,constant region 3C23K antibody, light chain, SEQ ID NO: 7 SEQ ID NO: 34variable region, without leader 3C23K antibody, light chain, SEQ ID NO:8 SEQ ID NO: 36 variable region, with leader 3C23K antibody, heavychain, SEQ ID NO: 9 SEQ ID NO: 38 variable region, without leader 3C23Kantibody, heavy chain, SEQ ID NO: 10 SEQ ID NO: 40 variable region, withleader 3C23K antibody, light chain, SEQ ID NO: 11 SEQ ID NO: 82 withoutleader 3C23K antibody, light chain, SEQ ID NO: 12 SEQ ID NO: 84 withleader 3C23K antibody, heavy chain, SEQ ID NO: 13 SEQ ID NO: 86 withoutleader 3C23K antibody, heavy chain, SEQ ID NO: 14 SEQ ID NO: 88 withleader

Even more particularly, the present invention relates to apharmaceutical composition comprising, as active ingredient, incombination with a pharmaceutically acceptable vehicle, an anticanceragent, and a monoclonal antibody binding AMHR-II, in which said mutatedhumanized monoclonal antibody comprises or is constituted by:

a) a light chain comprising or constituted by:

-   -   a variable region the amino acid sequence of which is        represented by SEQ ID NO: 7 or SEQ ID NO: 8,    -   a constant region the amino acid sequence of which is        represented by SEQ ID NO: 3        b) a heavy chain comprising or constituted by:    -   a variable region the amino acid sequence of which is        represented by SEQ ID NO: 9 or SEQ ID NO: 10    -   a constant region the amino acid sequence of which is        represented by SEQ ID NO: 6

Even more particularly, the present invention relates to apharmaceutical composition comprising, as active ingredient, incombination with a pharmaceutically acceptable vehicle, an anticanceragent, and a monoclonal antibody binding AMHR-II, in which said mutatedhumanized monoclonal antibody comprises or is constituted by:

a) a light chain constituted by the amino acid sequence represented bySEQ ID NO: 11 (without leader) or SEQ ID NO: 12 (with leader), andb) a heavy chain comprising or constituted by the amino acid sequencerepresented by SEQ ID NO: 13 (without leader), or SEQ ID NO: 14 (withleader).

Even more particularly, the present invention relates to apharmaceutical composition comprising, as active ingredient, incombination with a pharmaceutically acceptable vehicle, an anticanceragent, and a monoclonal antibody binding AMHR-II, in which said mutatedhumanized monoclonal antibody is produced by the 3C23K clone.

The mutated humanized monoclonal antibody produced by the 3C23K clone isdescribed in detail in international application WO 2011/141653. Byreference to the humanized 12G4 monoclonal antibody, the 3C23K antibodyhas the mutations of the 3C_(—)23 antibody, as well as an additionalmutation, in the CDR of the variable region of the light chain (E184K)in which a glutamic acid is replaced with a lysine, i.e. replacement ofan acidic amino acid with a basic amino acid consequently having atotally different charge since it is of opposite sign, yet stilldisplays an activity but especially an affinity far better than that ofthe unmutated humanized 12G4 antibody, and greater than that of theunmutated chimeric 12G4 antibody, and does not cause an immune reaction.

The mutated humanized monoclonal antibody produced by the 3C23K clonedisplays fucosylation with a content of about 50%.

According to another aspect, the present invention relates to apharmaceutical composition comprising, as active ingredient, incombination with a pharmaceutically acceptable vehicle, an anticanceragent, and a monoclonal antibody binding AMHR-II, in which said mutatedhumanized monoclonal antibody is produced by a clone described inapplication WO 2011/141653 and selected from the group consisting of:3C_(—)23, 6B_(—)78, 4C_(—)35 and 5B_(—)42.

In a particular embodiment, the invention relates to a pharmaceuticalcomposition comprising, as active ingredient, in combination with apharmaceutically acceptable vehicle, an anticancer agent, and anantibody binding AMHR-II, in which said antibody is a recombinantantibody produced by animal transgenesis.

This recombinant antibody may thus be produced by any technique known toa person skilled in the art, for example by recombination in a hostcell, transformed with one or more vector(s) that allow the expressionand/or the secretion of the nucleotide sequences coding for the heavychain and/or the light chain of the antibody. The vector generallycomprises a promoter, translation start and stop signals, as well asappropriate regions for the regulation of transcription. It ismaintained stably in the host cell and may optionally possess particularsignals that specify the secretion of the translated protein. Thesedifferent elements are selected and optimized by a person skilled in theart in relation to the cellular host used. Such vectors are prepared bymethods commonly used by a person skilled in the art, and the resultantclones may be introduced into a suitable host by standard methods. Thecellular host may be selected from prokaryotic or eukaryotic systems,for example bacterial cells but also yeast cells or animal cells,especially mammalian cells. The mammalian cells preferred for theproduction of the monoclonal antibodies are the YB2/0 rat line, the CHOhamster line, PER.C6™ (Crucell), 293, K562, NSO, SP2/0, BHK or COS. Itis also possible to use insect cells. Another method of production isthe expression of the recombinant antibody in transgenic organisms, forexample in plants or especially in the milk of transgenic animals suchas the rabbit, goat or pig. According to a preferred embodiment, theantibody is produced in the milk of non-human transgenic mammals,genetically modified to produce this glycoprotein. Preferably, it ismilk of a transgenic rabbit or goat, preferably in the milk of atransgenic goat. Advantageously, the antibody produced by animaltransgenesis, in particular in the mammary glands of a transgenic goat,has a glycosylation with a high degree of galactosylation, for examplegreater than 70%.

In particular, the invention relates to a pharmaceutical compositioncomprising, as active ingredient, in combination with a pharmaceuticallyacceptable vehicle, an anticancer agent, and an antibody bindingAMHR-II, in which the anticancer agent is paclitaxel or a platinum saltselected from the group consisting of: oxaliplatin, cisplatin andcarboplatin.

The anticancer agent may also be selected from chemotherapeutic agentsother than the platinum salts, small molecules, monoclonal antibodies orelse anti-angiogenesis peptibodies.

The chemotherapeutic agents other than the platinum salts include theintercalating agents (blocking of DNA replication and transcription),such as the anthracyclines (doxorubicin, pegylated liposomaldoxorubicin), the topoisomerase inhibitors (camptothecin andderivatives: Karenitecin, topotecan, irinotecan), or else SJG-136, theinhibitors of histone deacetylase (vorinostat, belinostat, valproicacid), the alkylating agents (bendamustine, glufosfamide, temozolomide),the anti-mitotic plant alkaloids, such as the taxanes (docetaxel,paclitaxel), the vinca alkaloids (vinorelbine), the epothilones(ZK-Epothilone, ixabepilone), the anti-metabolites (gemcitabine,elacytarabine, capecitabine), the kinesin spindle protein (KSP)inhibitors (ispinesib), trabectedin or else ombrabulin (combretastatinA-4 derivative).

Among the small molecules there are the poly(ADP-ribose)polymerase(PARP) inhibitors: olaparib, iniparib, veliparib, rucaparib, CEP-9722,MK-4827, BMN-673, the kinase inhibitors, such as the tyrosine kinaseinhibitors (TKI) among which there may be mentioned the anti-VEGFRmolecules (sorafenib, sunitinib, cediranib, vandetanib, pazopanib, BIBF1120, semaxanib, Cabozantinib, motesanib), the anti-HER2/EGFR molecules(erlotinib, gefitinib, lapatinib), the anti-PDGFR molecules (imatinib,BIBF 1120), the anti-FGFR molecules (BIBF 1120), the aurorakinase/tyrosine kinase inhibitors (ENMD-2076), the Src/Abl kinaseinhibitor (Saracatinib), or also Perifosine, Temsirolimus (mTORinhibitor), alvocidib (cyclin-dependent kinase inhibitor), Volasertib(inhibitor of PLK1 (polo-like kinase 1) protein, LY2606368 (inhibitor ofcheckpoint kinase 1 (chk 1), GDC-0449 (Hedgehog Pathway Inhibitor),Zibotentan (antagonist of the ETA-receptor), Bortezomib, Carfilzomib(proteasome inhibitor), cytokines such as IL-12, IL-18, IL-21,INF-alpha, INF-gamma.

Among the antibodies, there may be mentioned, the anti-VEGF:bevacizumab, the anti-VEGFR: ramucirumab, the anti-HER2/EGFRs:trastuzumab, pertuzumab, cetuximab, panitumumab, MGAH22, matuzumab,anti-PDGFR alpha: IMC-3G3, the anti-folate receptor: farletuzumab, theanti-CD27: CDX-1127, the anti-CD56: BB-10901, the anti-CD105: TRC105,the anti-CD276: MGA271, the anti-AGS-8: AGS-8M4, the anti-DRS: TRA-8,the anti-HB-EGF: KHK2866, the anti-mesothelins: amatuximab, BAY 94-9343(immunotoxin), catumaxomab (EpCAM/CD3 bispecific antibody), theanti-IL2R: daclizumab, the anti-IGF-1R: ganitumab, the anti-CTLA-4:ipilimumab, the anti-Lewis Y: Hu3S193, SGN-15 (immunotoxin), theanti-CAl25: oregovomab, the anti-HGF: rilotumumab, the anti-IL6:siltuximab, the anti-TR2: tigatuzumab, the anti-alpha5 beta1 integrin:volociximab, the anti-HB-EGF: KHK2866.

The anti-angiogenesis peptibodies are selected from AMG 386 and CVX-241.

More particularly, the invention relates to a pharmaceutical compositioncomprising, as active ingredient, in combination with a pharmaceuticallyacceptable vehicle, an anticancer agent, and an antibody bindingAMHR-II, in which the anticancer agent is carboplatin.

Even more particularly, the invention relates to a pharmaceuticalcomposition comprising, as active ingredient, in combination with apharmaceutically acceptable vehicle, an anticancer agent, and anantibody binding AMHR-II, in which the mutated humanized monoclonalantibody is produced by the 3C23K clone and the anticancer agent iscarboplatin.

In a particular aspect, the invention relates to a pharmaceuticalcomposition comprising, as active ingredient, in combination with apharmaceutically acceptable vehicle, an anticancer agent, and anantibody binding AMHR-II, in which the anticancer agent is paclitaxel.

In a more particular aspect, the invention relates to a pharmaceuticalcomposition comprising, as active ingredient, in combination with apharmaceutically acceptable vehicle, an anticancer agent, and anantibody binding AMHR-II, in which the mutated humanized monoclonalantibody is produced by the 3C23K clone and the anticancer agent ispaclitaxel.

In a particular aspect, the present invention relates to apharmaceutical composition comprising, as active ingredient, incombination with a pharmaceutically acceptable vehicle, an anticanceragent, and an antibody binding AMHR-II, in a formulation intended foradministration by the intravenous or intraperitoneal route.

The pharmaceutical composition of the invention may be administered byany suitable administration route, for example by the parenteral, oral,sublingual, vaginal, rectal, or transdermal route, preferably byintravenous, subcutaneous or intradermal injection. Intramuscular,intraperitoneal, intrasynovial, intrathecal or intratumoral injection isalso possible. The injections may be carried out in the form of a bolus,or by continuous infusion. When the antibody composition and thecomposition of anticancer agent are administered separately, thesecompositions may be in an identical or different form of administration.

The preparations for parenteral administration may include sterileaqueous or non-aqueous solutions, suspensions or emulsions. Examples ofnon-aqueous solvents are propylene glycol, polyethylene glycol,vegetable oils, such as olive oil, or injectable organic esters such asethyl oleate. Aqueous vehicles comprise water, alcohol/water solutions,and emulsions or suspensions.

The pharmaceutical compositions according to the inventionadvantageously comprise one or more pharmaceutically acceptableexcipients or vehicles. There may be mentioned for example saline,physiological, isotonic, buffered solutions, etc., compatible withpharmaceutical use and known to a person skilled in the art. Thecompositions may contain one or more agents or vehicles selected fromdispersants, solubilizers, stabilizers, preservatives, etc. Agents orvehicles usable in formulations (liquid and/or injectable and/or solid)are in particular methylcellulose, hydroxymethylcellulose,carboxymethylcellulose, polysorbate 80, mannitol, gelatin, lactose,vegetable oils, acacia, etc. The compositions may be formulated in theform of injectable suspensions, gels, oils, tablets, suppositories,powders, hard gelatine capsules, soft capsules, etc.

According to a particular aspect, the invention relates to apharmaceutical composition comprising, as active ingredient, incombination with a pharmaceutically acceptable vehicle, an anticanceragent and an antibody binding AMHR-II, in which the therapeuticallyeffective quantity of antibody administered to a patient is in a rangefrom about 0.07 mg to about 35000 mg, preferably from about 0.7 mg toabout 7000 mg, preferably from about 0.7 mg to about 1400 mg, preferablyfrom about 0.7 mg to about 700 mg, and more preferably from about 0.7 mgto about 70 mg.

The dosage of the active ingredient depends in particular on theadministration method, and is easily determined by a person skilled inthe art. A therapeutically effective quantity (unit dose) of antibodymay vary from 0.01 mg/kg to 500 mg/kg, preferably from 0.1 mg/kg to 500mg/kg, preferably from 0.1 mg/kg to 100 mg/kg, preferably from 0.1 mg/kgto 20 mg/kg, preferably from 0.1 mg/kg to 10 mg/kg, and more preferablyfrom 1 mg/kg to 10 mg/kg, in one or more weekly administrations, forseveral weeks or months. The effective unit dose may therefore easily bededuced from a dose calculated for an “average” patient with a weight of70 kg.

According to another particular aspect, the invention relates to apharmaceutical composition comprising, as active ingredient, incombination with a pharmaceutically acceptable vehicle, an anticanceragent and an antibody binding AMHR-II, in which the therapeuticallyeffective quantity of anticancer agent administered to a patient is in arange from about 10 mg to about 700 mg, preferably in a range from about20 mg to about 350 mg, and preferably is about 110 mg.

The dosage of the anticancer agent depends in particular on theadministration method, and is easily determined by a person skilled inthe art. A therapeutically effective quantity (unit dose) may vary from0.2 mg/m² to 10 g/m², preferably from 0.2 mg/m² to 1 g/m², preferablyfrom 2 mg/m² to 1 g/m², preferably from 20 mg/m² to 1 g/m², and morepreferably from 20 mg/m² to 0.5 g/m², in one or more weeklyadministrations, for several weeks or months. The effective unit dosemay therefore be deduced from a dose calculated for an “average” patientwhose body surface area is about 1.8 m².

According to an even more particular aspect, the invention relates to apharmaceutical composition comprising, as active ingredient, incombination with a pharmaceutically acceptable vehicle, an anticanceragent and an antibody binding AMHR-II, in which the therapeuticallyeffective quantity of anticancer agent administered to a patient isabout 110 mg, and the therapeutically effective quantity of antibodyadministered to the patient is about 70 mg.

The invention also relates to a composition comprising an anticanceragent and an antibody binding the human anti-Müllerian hormone type IIreceptor (AMHR-II), for use as a medicinal product in the prevention ortreatment of a pathology associated with the human anti-Müllerianhormone type II receptor (AMHR-II).

By “treatment” is meant the means for treating a manifest pathology, thesymptoms of which are visible. By “prevention” is meant the means forpreventing said pathology from occurring.

A pathology associated with the human anti-Müllerian hormone type IIreceptor (AMHR-II) may in particular be:

-   -   ovarian cancer, in particular metastatic ovarian cancer, and its        various subtypes, in particular: serous, clear-cell,        endometrioid, mucinous,    -   germ cell cancer,    -   endometrial cancer,    -   mixed Müllerian malignant tumour of the uterus,    -   leiomyosarcoma,    -   endometrial stromal sarcoma,    -   prostate cancer,    -   testicular cancer.

Tumours expressing the AMHR-II antigen are targeted preferentially, i.e.tumours in which a significant level of expression of the AMHR-IIantigen in a cell is observed, preferably on the surface of the cells.

According to the invention, the two therapeutic agents are used incombination in order to potentiate the antiproliferative effects of bothof them.

In a particular aspect, the invention relates to a composition for useas a medicinal product in the prevention or treatment of a pathologyassociated with AMHR-II, comprising an anticancer agent and an antibodybinding AMHR-II, in which said antibody is a polyclonal antibody.

According to another particular aspect, the invention relates to acomposition for use as a medicinal product in the prevention ortreatment of a pathology associated with AMHR-II, comprising ananticancer agent and an antibody binding AMHR-II, in which said antibodyis a monoclonal antibody, and preferably a chimeric or humanized 12G4monoclonal antibody.

According to another particular aspect, the invention relates to acomposition for use as a medicinal product in the prevention ortreatment of a pathology associated with AMHR-II, comprising ananticancer agent and an antibody binding AMHR-II, in which said antibodyis a mutated humanized 12G4 antibody, or a fragment of mutated humanized12G4 monoclonal antibody, in which said monoclonal antibody comprises atleast one mutation in the light and/or heavy chain.

According to another particular aspect, the invention relates to acomposition for use as a medicinal product in the prevention ortreatment of a pathology associated with AMHR-II, comprising ananticancer agent and an antibody binding AMHR-II, in which said antibodyis a mutated humanized 12G4 antibody, or a fragment of mutated humanized12G4 monoclonal antibody, in which said monoclonal antibody comprises atleast one mutation in the light and/or heavy chain and has an AMHR-IIaffinity characterized by a K_(D) preferably less than 10⁻⁷ M, inparticular less than 10⁻⁸ M, in particular in the range from 10⁻⁹ M to10⁻¹¹ M. According to another particular aspect, the invention relatesto a composition for use as a medicinal product in the prevention ortreatment of a pathology associated with AMHR-II, comprising ananticancer agent and an antibody binding AMHR-II, in which said antibodyis a mutated humanized 12G4 antibody, or a fragment of mutated humanized12G4 monoclonal antibody, in which said monoclonal antibody comprises atleast one mutation in the light and/or heavy chain.

According to another particular aspect, the invention relates to acomposition for use as a medicinal product in the prevention ortreatment of a pathology associated with AMHR-II, comprising ananticancer agent and an antibody binding AMHR-II, in which said 12G4antibody comprises or is constituted by:

-   -   a) a light chain comprising or constituted by:        -   a variable region the amino acid sequence of which is            represented by SEQ ID NO: 1 or SEQ ID NO: 2, and        -   a constant region the amino acid sequence of which is            represented by SEQ ID NO: 3 or having at least 80% homology            with SEQ ID NO: 3,    -   b) a heavy chain comprising or constituted by:        -   a variable region the amino acid sequence of which is            represented by SEQ ID NO: 4, or SEQ ID NO: 5, and        -   a constant region the amino acid sequence of which is            represented by SEQ ID NO: 6 or by a sequence having at least            80% homology with SEQ ID NO: 6,            in which the humanized 12G4 monoclonal antibody is mutated,            comprises at least one mutation in the light and/or heavy            chain, and has a K_(D) for the human anti-Müllerian hormone            type II receptor (AMHR-II) preferably less than 10⁻⁷ M, in            particular less than 10⁻⁸ M, in particular in the range from            10⁻⁹ M to 10⁻¹¹ M.

In a particular aspect, the invention relates to a composition for useas a medicinal product in the prevention or treatment of a pathologyassociated with AMHR-II, comprising an anticancer agent and an antibodybinding AMHR-II, in which the humanized monoclonal antibody comprises oris constituted by

-   -   a) a light chain comprising or constituted by:        -   a variable region the amino acid sequence of which is            represented by SEQ ID NO: 7 or SEQ ID NO: 8,        -   a constant region the amino acid sequence of which is            represented by SEQ ID NO: 3    -   b) a heavy chain comprising or constituted by:        -   a variable region the amino acid sequence of which is            represented by SEQ ID NO: 9 or SEQ ID NO: 10, and        -   a constant region the amino acid sequence of which is            represented by SEQ ID NO: 6.

In a particular aspect, the invention relates to a composition for useas a medicinal product in the prevention or treatment of a pathologyassociated with AMHR-II, comprising an anticancer agent and an antibodybinding AMHR-II, in which the humanized 12G4 monoclonal antibody isproduced by the clone 3C-23K.

In a particular aspect, the invention relates to a composition for useas a medicinal product in the prevention or treatment of a pathologyassociated with AMHR-II, comprising an anticancer agent and an antibodybinding AMHR-II, in which the humanized 12G4 monoclonal antibody is afragment of the humanized 12G4 monoclonal antibody produced by the clone3C-23K.

In a particular aspect, the invention relates to a composition for useas a medicinal product in the prevention or treatment of a pathologyassociated with AMHR-II, comprising an anticancer agent and an antibodybinding AMHR-II, in which the antibody is a recombinant antibodyproduced by animal transgenesis.

In a particular aspect, the invention relates to a composition for useas a medicinal product in the prevention or treatment of a pathologyassociated with AMHR-II, comprising an anticancer agent and an antibodybinding AMHR-II, in which the pathology associated with the humananti-Müllerian hormone type II receptor (AMHR-II) is cancer, andparticularly ovarian cancer.

In a particular aspect, the invention relates to a composition for useas a medicinal product in the prevention or treatment of a pathologyassociated with AMHR-II, comprising an anticancer agent and an antibodybinding AMHR-II, in which the anticancer agent is paclitaxel or aplatinum salt selected from the group constituted by: oxaliplatin,cisplatin, carboplatin.

In a particular aspect, the invention relates to a composition for useas a medicinal product in the prevention or treatment of a pathologyassociated with AMHR-II, comprising an anticancer agent and an antibodybinding AMHR-II, in which the anticancer agent is carboplatin.

In a particular aspect, the invention relates to a composition for useas a medicinal product in the prevention or treatment of a pathologyassociated with AMHR-II, comprising an anticancer agent and an antibodybinding AMHR-II, in which the anticancer agent is paclitaxel.

In a particular aspect, the invention relates to a composition for useas a medicinal product in the prevention or treatment of a pathologyassociated with AMHR-II, comprising the 12G4 monoclonal antibodyproduced by the 3C23K clone and carboplatin.

In another particular aspect, the invention relates to a composition foruse as a medicinal product in the prevention or treatment of a pathologyassociated with AMHR-II, comprising the 12G4 monoclonal antibodyproduced by the 3C23K clone and paclitaxel.

In another particular aspect, the invention relates to a composition foruse as a medicinal product in the prevention or treatment of a pathologyassociated with AMHR-II, comprising an anticancer agent and an antibodybinding AMHR-II, in a formulation intended for administration by theintravenous or intraperitoneal route.

In another particular aspect, the invention relates to a composition foruse as a medicinal product in the prevention or treatment of a pathologyassociated with AMHR-II, comprising an anticancer agent and an antibodybinding AMHR-II, the monoclonal antibody and the anticancer agent beingintended for separate, simultaneous or sequential administration.

The antibody and the anticancer agent may be combined within one and thesame pharmaceutical composition, or may be used in the form of separatepharmaceutical compositions, which may be administered simultaneously orsequentially. In particular, the products may be administeredseparately, namely either concomitantly, or independently, for examplewith a time gap.

More particularly, the invention relates to a composition for use as amedicinal product in the prevention or treatment of a pathologyassociated with AMHR-II, comprising an anticancer agent and an antibodybinding AMHR-II, in which the antibody and the anticancer agent arecombined within the same pharmaceutical composition.

According to another particular aspect, the invention relates to acomposition for use as a medicinal product in the prevention ortreatment of a pathology associated with AMHR-II, comprising ananticancer agent and an antibody binding AMHR-II, in which thetherapeutically effective quantity of antibody administered to a patientis in a range from about 0.07 mg to about 35 000 mg, preferably fromabout 0.7 mg to about 7000 mg, preferably from about 0.7 mg to about1400 mg, preferably from about 0.7 mg to about 700 mg, and morepreferably from about 0.7 mg to about 70 mg.

According to another particular aspect, the invention relates to acomposition for use as a medicinal product in the prevention ortreatment of a pathology associated with AMHR-II, comprising ananticancer agent and an antibody binding AMHR-II, in which thetherapeutically effective quantity of anticancer agent administered to apatient is in a range from about 10 mg to about 700 mg, preferably in arange from about 20 mg to about 350 mg, and preferably about 110 mg.

According to another particular aspect, the invention relates to acomposition for use as a medicinal product in the prevention ortreatment of a pathology associated with AMHR-II, comprising ananticancer agent and an antibody binding AMHR-II, in which thetherapeutically effective quantity of antibody administered to a patientis about 70 mg and the dose of anticancer agent administered to thepatient is about 110 mg.

In a preferred embodiment, the dosage of anticancer agent, in particularcarboplatin or paclitaxel, is in a range from about 0.01 mg/kg to about500 mg/kg, for example 0.1 mg/kg to 300 mg/kg, or from about 0.1 mg to20 g per day.

As a variant, a higher initial loading dose, followed by one or morelower doses may also be administered. In another variant, an initialloading dose that is not so high, followed by one or more higher dosesmay also be administered.

In a particular embodiment, the anti-AMHR-II antibody and carboplatinmay be used in an antibody/carboplatin ratio in a range from about 10/1to about 0.1/1, in particular from about 10/1 to about 1/1, or fromabout 1/1 to about 0.1/1.

In another particular embodiment, the anti-AMHR-II antibody andpaclitaxel may be used in an antibody/paclitaxel ratio in a range fromabout 10/1 to about 0.1/1, in particular from about 10/1 to about 1/1,or from about 1/1 to about 0.1/1.

The invention further relates to a product comprising an antibodybinding the human anti-Müllerian hormone type II receptor (AMHR-II) andan anticancer agent, in the form of a combined preparation, forsimultaneous, sequential or separate use as a medicinal product intendedfor preventing or treating a pathology associated with the humananti-Müllerian hormone type II receptor (AMHR-II), in particular cancer,and more particularly ovarian cancer.

According to a particular aspect, the invention relates to a productcomprising an antibody binding the human anti-Müllerian hormone type IIreceptor (AMHR-II) and an anticancer agent, in the form of a combinedpreparation, for simultaneous, sequential or separate use as a medicinalproduct intended for preventing or treating a pathology associated withthe human anti-Müllerian hormone type II receptor (AMHR-II), inparticular cancer, and more particularly ovarian cancer, forsimultaneous use of the antibody and of the anticancer agent.

According to another particular aspect, the invention relates to aproduct comprising an antibody binding the human anti-Müllerian hormonetype II receptor (AMHR-II) and an anticancer agent, in the form of acombined preparation, for simultaneous, sequential or separate use as amedicinal product intended for preventing or treating a pathologyassociated with the human anti-Müllerian hormone type II receptor(AMHR-II), in particular cancer, and more particularly ovarian cancer,for sequential use of the antibody and of the anticancer agent, in whichthe antibody is administered prior to the anticancer agent.

According to another particular aspect, the invention relates to aproduct comprising an antibody binding the human anti-Müllerian hormonetype II receptor (AMHR-II) and an anticancer agent, in the form of acombined preparation, for simultaneous, sequential or separate use as amedicinal product intended for preventing or treating a pathologyassociated with the human anti-Müllerian hormone type II receptor(AMHR-II), in particular cancer, and more particularly ovarian cancer,for sequential use of the antibody and of the anticancer agent, in whichthe anticancer agent is administered prior to the antibody.

The following figures, tables and examples illustrate the invention,without limiting its scope.

FIG. 1A shows the variation of the mean tumour volumes, expressed inmm³, on the y-axis, as a function of the number of days counting fromthe day of injection of the tumour cells. FIG. 1B shows the curve of themedian tumour volumes in mm³, on the y-axis, as a function of the numberof days counting from the day of injection of the tumour cells. In eachof the graphs, the curve joining the filled squares represents the meanvalue of the mice in the control group, the continuous curve joining thediamonds represents the group of mice treated with the irrelevantantibody LFB-R565, the curve joining the filled triangles represents thegroup of mice treated with the 3C23K antibody, the continuous curvejoining the filled circles represents the group of mice treated withcarboplatin, the dotted curve joining the filled circles represents thegroup of mice treated with carboplatin and with the irrelevant antibodyLFB-R565, the dotted curve joining the filled triangles represents thegroup of mice treated with carboplatin and with the 3C23K antibody.

FIGS. 2A to 2F show the curves of the individual tumour volumes, bygroups. For each of the graphs 2A to 2F, the x-axis shows the number ofdays counting from the day of injection of the tumour cells, and they-axis shows the tumour volume, each of the curves show the evolution oftumour volume for one animal. FIG. 2A shows the control group of mice,FIG. 2B shows the group of mice treated with the irrelevant antibodyLFB-R565, FIG. 2C shows the group of mice treated with the 3C23Kantibody, FIG. 2D shows the group of mice treated with carboplatin, FIG.2E shows the group of mice treated with carboplatin and with theirrelevant antibody, FIG. 2F shows the group of mice treated withcarboplatin and with the 3C23K antibody.

FIGS. 3A, 3B and 3C show the variation of the mean tumour volumes (3A)or the median tumour volumes (3B) and percentage survival, in 4 groupsof mice treated respectively with the 3C23K antibody in monotherapy,paclitaxel in monotherapy or the combination of the 3C23K antibody andpaclitaxel. A solution of NaCl was used as the control. FIG. 3A showsthe variation of the mean the tumour volumes, expressed in mm³, on they-axis, as a function of the number of days counting from the day ofinjection of the tumour cells. FIG. 3B shows the curve of the mediantumour volumes in mm³, on the y-axis, as a function of the number ofdays counting from the day of injection of the tumour cells. FIG. 3Cshows the respective percentage survival for the four groups, on they-axis, as a function of the number of days counting from the day ofinjection of the tumour cells. In FIGS. 3A to 3C: the curves joining thediamonds correspond to the group treated with the control solution; thecurves joining the squares correspond to the group treated with the3C23K antibody in monotherapy; the curves joining the round dotscorrespond to the group treated with paclitaxel; the curves joining thetriangles correspond to the group treated with the combination of the3C23K antibody and paclitaxel.

FIGS. 4A, 4B, 4C and 4D show respectively the curve of the individualtumour volumes in 4 groups of mice treated respectively with the controlsolution (4A), the 3C23K antibody in monotherapy (4B), paclitaxel inmonotherapy (4C), or the combination of the 3C23K antibody andpaclitaxel (4D).

Table 1 gives a summary of the treatment schedule.

Table 2 gives a summary of the raw data for individual tumour volume.

Table 3 gives a summary of the raw data for mean tumour volume andstandard deviation (SD).

Table 4 gives the raw data for the median tumour volumes and T/C ratio.

Table 5 gives a statistical analysis of the tumour volumes.

Table 6 gives a summary of the raw data for individual body weight.

Table 7 gives a summary of the raw data for mean body weight andstandard deviation (SD).

Table 8 gives a summary of the raw data for individual body weight.

Table 9 gives a summary of the raw data for individual tumour volume.

Table 10 gives a summary of the raw data for the change in mean bodyweight.

Table 11 gives a summary of the raw data for the mean tumour volumes.

Table 12 gives a summary of the raw data for the median tumour volumesand for the T/C ratios.

Table 13 gives the results of statistical analysis.

Table 14 gives a summary of the raw data for the survival parameter

Table 15 gives the survival parameters

EXAMPLE Evaluation of the Activity of the Antibody Produced by the 3C23KClone (3C23K Antibody), in Monotherapy or in Combination withCarboplatin, in a Model of Human Ovarian Cancer Cov434-AMHRII Asc1a5 inthe Female Swiss Nude Mouse

1. Protocol

Female Swiss nude mice (Harlan Laboratories) were injectedsubcutaneously (s.c.) with 7.10⁶ cells of Cov434-AMHRII Asc1a5 (cellline of human ovarian cancer transfected with cDNA AMHRII) in Matrigel(1:1 ratio) under a volume of 150 μL on day 0 (D0).

The 3C23K antibody was evaluated according to the following scheme: 2times per week for 6 weeks, for a total of 12 injections at about 10mg/kg/injection, said administration regimen being designated below as“Q3-4D12”. Another group of mice was treated with an irrelevant antibodyLFB-R565, administered at about 10 mg/kg/injection, according to theregimen Q3-4D12.

Carboplatin was evaluated, at a sub-optimum dose, i.e. about 60mg/kg/injection, according to the following scheme: once a week, for 4weeks, said administration regimen being designated below as “Q7D4”.

Carboplatin was also evaluated in combination with the 3C23K antibody orwith the irrelevant antibody LFB-R565. Carboplatin was administered atabout 60 mg/kg/injection according to the Q7D4 regimen and the 3C23Kantibody or the irrelevant antibody LFB-R565 at about 10 mg/kg/injectionaccording to the regimen Q3-4D12.

The mice were randomized on D11, when the volume of the tumours wasbetween 50 and 158 mm³, and the treatments began on D13 (9 mice pergroup). The description of the treatment dates is presented in Table 1.

In the context of evaluating the activity of the combination of the3C23K antibody and paclitaxel relative to that of the 3C23K antibody inmonotherapy or of paclitaxel in monotherapy, the 3C23K antibody wasinjected according to the following regimen: once weekly for 4 weeks,for a total of 4 injections, at a dose of about 10 mg/kg/injectionaccording to the Q7D4 regimen; paclitaxel was injected according to thefollowing regimen: once weekly for 4 weeks, for a total of 4 injections,at a dose of about 15 mg/kg/injection (Q7D4 regimen).

The mice were randomized on D13, when the volume of the tumours wasbetween 58 and 150 mm³, and the treatments began on D14 (8 mice pergroup).

a. Monitoring the Experiments In Vivo

The tumours were usually measured twice weekly. Tumour volume (TV) wascalculated using the following formula, in which the length correspondsto the largest of the tumour diameters, the width corresponds to thesmallest of the tumour diameters and the tumour height: TV(mm³)=(length×width×height)/2.

The curves of the individual volume of the tumours were plotted.

In addition, for each group, curves of tumour growth were plotted usingthe calculated mean tumour volumes or the median tumour volumes.

The animals were sacrificed when the tumour volumes reached 2000 mm³ orfor ethical reasons. The curves of mean values and median values as wellas the statistical analyses were stopped when 20% of the mice in thegroup were dead.

In one experiment with 9 mice per group, the curves and analyses weretherefore stopped when fewer than 8 values per group had been obtained(8 mice alive).

In one experiment with 8 mice per group, the curves and analyses werestopped when fewer than 7 values per group had been obtained (7 micealive).

b. Evaluation of the Efficacy of the Treatment

Inhibition of tumour growth, defined as the ratio between the mediantumour volumes of the treated mice relative to the treated controlgroups (T/C) was calculated as follows:

T/C=(mean TV of the treated group/mean TV of the control group)×100

The National Cancer Institute used the following criteria for evaluatingthe anti-tumour activity of a product (Bissery et al., 1991):

T/C greater than 42%, the product is considered to be ineffective

T/C between 42% and 10%, the product displays an anti-tumour effect

T/C less than 10%, the product is really effective.

Moreover, to identify whether the treatment has a toxic effect, theweight of the mice was monitored individually once a week. The mean bodyweight of the mice was calculated for each group, until 20% of the micein the group were dead.

c. Statistical Analyses

The statistical differences between the different groups were analysedby ANOVA comparison using the Statgraphics centurion XV software.

The ANOVA table breaks down the variance into two components: aninter-group component and an intra-group component. The ratio F is theratio of the inter-group estimate to the intra-group estimate. When theP value of test F is greater than or equal to 0.05, there is nostatistically significant difference between the mean values of the twogroups, with a confidence level of 95%. P values less than 0.05 indicatea significant difference between the mean values of the two groups witha confidence level of 95%.

The raw data of the statistical analyses in experiments with the F ratioand P value are presented in the appendix for all the experiments.

A Kruskal-Wallis test was also carried out. When the P value is greaterthan or equal to 0.05, there is no statistically significant differencebetween the median of the two groups, with a confidence level of 95%. Pvalues less than 0.05 indicate a significant difference between themedians of the two groups with a confidence level of 95%.

2. Results

2.1 Evaluation of Anti-Tumour Activity of the 3C23K Antibody inCombinations with Carboplatin

It appears that the 3C23K antibody or 3C23K displays significantanti-tumour activity compared to the control (NaCl solution) and to theLFB-R565 antibody (or LFB-R565) (FIGS. 1 and 2; Tables 2, 3, 4 and 5).Relative to the control, the 3C23K T/C ratios decreased from D14 to D36,on D18 the T/C ratio was 33% and was constantly less than 23% up to D36(Table 4). Relative to the irrelevant antibody LFB-R565, the T/C ratiosdecreased from D14 to D36, on D18 the T/C ratio was 41% and wasconstantly less than 26% up to D36 (Table 4).

Moreover, the treatment with the irrelevant antibody LFB-R565 does notdisplay anti-tumour activity relative to the control (FIGS. 1 and 2;Tables 2, 3, 4 and 5) since the T/C ratios were never less than 68%(Table 4).

Carboplatin also displays significant anti-tumour activity compared tothe control or to the irrelevant antibody LFB-R565 (FIGS. 1 and 2;Tables 2, 3, 4 and 5). From D18 to D36, the T/C ratios relative to thecontrol decreased gradually and the value was 40% on D36, indicating ananti-tumour activity (Table 4). Similarly, when compared to a grouptreated with the irrelevant antibody LFB-R565, the T/C ratio decreasedfrom D25 to D36, reaching 35% on D36 (Table 4).

Results similar to those for the group treated with carboplatin wereobtained with the combination carboplatin+LFB-R565, compared to thecontrol group. From D18 to D36, the T/C ratios decreased gradually and,on D36, the T/C ratio was 40%, demonstrating an anti-tumour activity(Table 4).

When the group treated with carboplatin+LFB-R565 was regarded as thetreated group and was compared to LFB-R565 alone, a decrease in the T/Cratios was observed starting from D22 and two T/C ratios less than 42%were found, on D25 and D36 (Table 4).

Moreover, there was no difference between the groups treated withcarboplatin+LFB-R565 or with carboplatin alone (FIG. 1; Tables 2, 3, 4and 5).

3C23K (10 mg/kg, Q3-4D12) and carboplatin (60 mg/kg, Q7D4) used inmonotherapy showed an anti-tumour effect in this Cov434-AMHRII Asc1a5xenografted tumour model at the doses tested (FIGS. 1 and 2; Tables 2,3, 4 and 5).

Moreover, in the test conditions, 3C23K showed an anti-tumour effectgreater than carboplatin. In fact, the T/C ratio of the group treatedwith 3C23K relative to the group treated with carboplatin decreased fromD14 to D39, and from D22 to D39 the T/C ratios were less than 42%, whichindicates an anti-tumour activity of 3C23K greater than that ofcarboplatin (Table 4).

When the group treated with carboplatin combined with 3C23K was comparedto a control group, the combination showed a very strong anti-tumoureffect: on D18 the T/C ratio was 24% and from D22 to D36 the T/C ratioswere less than 10% (FIGS. 1 and 2; Tables 2, 3, 4 and 5).

Moreover, the combination of carboplatin (60 mg/kg, Q7D4) and 3C23K (10mg/kg, Q3-4D12) showed an anti-tumour effect stronger than each of thecomponents, 3C23K (10 mg/kg, Q3-4D12) or carboplatin (60 mg/kg, Q7D4)used in monotherapy (FIGS. 1 and 2; Tables 2, 3, 4 and 5).

In fact, when the group treated with carboplatin+3C23K was compared withthe group treated with carboplatin, the combination showed a greateranti-tumour activity: on D14 the T/C ratio began to decrease and fromD25 to D43 the calculated ratio was less than 11%.

Moreover, when the group treated with 3C23K and carboplatin was comparedto the group treated with 3C23K, the combination showed an anti-tumouractivity greater than that of the monotherapy with 3C23K. On D18, theT/C ratio began to decrease, and was found to be less than 42% betweenD22 and D29, which indicates an anti-tumour advantage, and was found tobe less than 11% between D32 and D49, which indicates a largeanti-tumour advantage (FIGS. 1 and 2; Tables 2, 3, 4 and 5).

Moreover, the group treated with carboplatin and 3C23K also showed ananti-tumour advantage compared to carboplatin combined with anirrelevant antibody LFB-R565 (Table 4).

The treatments had no effect until day 32 (Tables 6 and 7). On D32, atransient decrease in body weight was observed for the groups treatedwith carboplatin, carboplatin+LFB-R565 and carboplatin+3C23K. Thisdecrease, after the third injection of carboplatin, on D28, was notgreater than 15%, in comparison with the previous measurements on D25.The decrease was similar for the groups treated with carboplatin orcarboplatin+LFB-R565 (about 15%) and was about 1% for the group treatedwith carboplatin+3C23K. In these three groups, the decrease in weight,slight and transient, was not regarded as a toxic effect as it was notconfirmed after the 4th injection of carboplatin, on D34 (Tables 6 and7).

3. Conclusions

In the present study the inventors evaluated the combination of 3C23K at10 mg/kg, Q3-4D12, and the sub-optimum dose of carboplatin 60 mg/kg,Q7D4, on female nude mouse models with the Cov434-AMHRII Asc1a5xenografted tumour, compared to a control group. An irrelevant antibody,LFB-R565 (10 mg/kg, Q3-4D12), was also tested, alone or in combinationwith carboplatin.

The results demonstrate that carboplatin (60 mg/kg Q7D4) exertsanti-tumour activity on Cov434-AMHRII Asc1a5. The results alsodemonstrate that 3C23K (10 mg/kg Q3-4D12) exerts anti-tumour activity onCov434-AMHRII Asc1a5.

However, the anti-tumour activity observed with carboplatin alone waslower than that observed with 3C23K alone (10 mg/kg, Q3-4D12).

The combination of 3C23K and carboplatin was demonstrated as displayingan advantage when compared to 3C23K alone or to carboplatin alone and asa minimum had an additive effect.

Finally, the anti-tumour activity of 3C23K alone or in combination withcarboplatin is specific, as no efficacy was observed with an irrelevantantibody, whether alone or in combination.

TABLE 1 Summary of the treatment schedule Treatment schedule Carboplatincontrol or Date Product vehicle LFB-R565 3C23K carboplatin carboplatin +LFB-R565 carboplatin + 3C23K 19-sep D11 Randomization of the animals20-sep D12 21-sep D13 TT TT TT TT TT 22-sep D14 TT TT TT 23-sep D15 TTTT TT TT TT 24-sep D16 25-sep D17 26-sep D18 TT TT TT TT TT 27-sep D1928-sep D20 29-sep D21 TT TT TT 30-sep D22 TT TT TT TT TT 01-oct D2302-oct D24 03-oct D25 TT TT TT TT TT 04-oct D26 05-oct D27 06-oct D28 TTTT TT 07-oct D29 TT TT TT TT TT 08-oct D30 09-oct D31 10-oct D32 TT TTTT TT TT 11-oct D33 12-oct D34 13-oct D35 TT TT TT 14-oct D36 TT TT TTTT TT 15-oct D37 16-oct D38 17-oct D39 TT TT TT TT TT 18-oct D40 19-octD41 20-oct D42 21-oct D43 TT TT TT TT TT 22-oct D44 23-oct D45 24-octD46 TT TT TT TT TT 25-oct D47 26-oct D48 27-oct D49

TABLE 2 Summary of the raw data for individual tumour volume Individualtumour volume Sep. 19, Sep. 22, Sep. 26, Sep. 30, Oct. 3, Oct. 7, Oct.10, Oct. 14, Oct. 17, Oct. 21, Oct. 24, Oct. 27, Mouse 2011 2011 20112011 2011 2011 2011 2011 2011 2011 2011 2011 Group number D11 D14 D18D22 D25 D29 D32 D36 D39 D43 D46 D49 NaCl 1-SM 91 126 250 504 819 9521020 2404 1-OD 72 169 143 297 303 1320 891 1260 1848 1564 2622 1-OG 96147 216 520 700 588 2993 1-2O 107 180 240 468 480 644 864 1540 1560 26251-2OD 99 192 144 216 330 462 660 1144 2280 2-SM 60 163 248 252 336 5041045 1617 2070 2-OD 74 120 135 392 441 936 1053 1589 2024 2-OG 158 234324 672 495 819 1181 1607 1386 2178 2-2O 77 123 108 180 1264 420 490 630828 912 1485 2080 LFB-R565 3-SM 90 203 173 378 432 792 1287 2321 3-OD 72110 140 126 216 792 356 588 720 995 1215 1672 3-OG 96 154 297 360 465840 1140 2166 2166 3-2O 153 175 460 324 520 588 690 1824 1790 2112 3-2OD108 154 192 363 504 980 1536 2052 4-SM 58 176 216 270 385 585 1122 19802576 4-OD 72 110 140 197 410 693 616 774 528 624 413 630 4-OG 81 95 108216 297 504 569 1584 1454 2079 4-2O 60 95 160 270 288 504 660 1144 10731672 1785 2321 3C23K 5-SM 98 104 72 60 0 0 0 0 0 0 0 0 5-OD 90 179 88150 126 322 501 653 784 1171 1683 1870 5-OG 61 60 36 30 0 27 27 87 210420 598 1188 5-2O 80 72 21 0 0 0 0 0 0 0 0 0 5-2OD 57 67 41 36 24 48 150 0 0 0 0 6-SM 147 216 140 131 180 225 275 363 315 631 504 784 6-OD 113166 80 77 74 146 180 182 182 393 275 655 6-OG 72 84 72 158 216 385 644784 1200 1403 1650 2646 6-2O 72 53 53 70 88 210 350 432 546 731 518 945carboplatin 7-SM 53 104 98 378 140 270 351 630 660 864 1071 1568 7-OD142 210 173 126 240 200 243 424 484 592 501 819 7-OG 61 162 112 360 270360 578 1148 1134 1881 2205 7-2O 102 210 336 324 616 768 864 1610 14952112 7-2OD 90 126 248 363 152 360 653 893 1488 2038 8-SM 72 123 140 270358 363 504 462 447 882 810 1125 8-OD 81 75 90 156 98 168 220 308 347438 378 480 8-OG 70 134 112 198 243 420 462 774 910 1105 1450 1650 8-2O114 168 90 168 210 187 275 594 423 672 878 980 LFB-R565 + 9-SM 116 203216 297 298 420 518 720 936 1323 1440 2112 carboplatin 9-OD 72 90 123126 112 180 385 594 608 900 965 1233 9-OG 63 90 126 112 91 144 180 350501 429 592 686 9-2O 102 150 180 220 170 330 336 726 564 675 570 9369-2OD 98 168 299 420 432 1040 1134 2288 10-SM 51 210 193 210 264 410 455704 774 1008 1378 1881 10-OD 130 180 338 280 193 865 798 1232 1449 17831320 2185 10-OG 68 72 85 135 123 192 266 462 840 1081 995 995 10-2O 8190 98 108 117 210 248 216 378 293 3C23K + 11-SM 63 168 140 120 105 96 8870 45 165 175 189 carboplatin 11-OD 123 102 54 38 24 40 18 0 24 66 69 9011-OG 83 95 45 24 23 35 45 48 50 11-2O 83 96 53 24 18 81 18 18 0 18 2324 11-2OD 123 112 112 24 0 0 0 18 0 18 34 81 12-SM 50 84 18 0 0 0 0 1811 24 23 28 12-OD 72 77 47 72 54 108 88 140 100 242 250 325 12-OG 107158 21 0 0 32 0 0 0 24 42 45 12-2O 65 60 72 98 132 112 105 160 184 357273 539

TABLE 3 Summary of the raw data for mean tumour volume and standarddeviation (SD) Mean tumour volume Day Group 11 14 18 22 25 29 32 36 3943 46 49 Control Mean 93 161 201 389 463 738 1133 1474 SD 10 13 25 58 67104 258 191 LFB-R565 Mean 10 14 38 31 37 58 140 222 SD 10 21 12 20 29 5184 105 146 181 233 321 3C23K Mean 88 111 67 79 79 151 221 278 360 528581 899 SD 10 21 12 20 29 51 84 105 146 181 233 321 carboplatin Mean 87146 155 260 258 344 461 760 821 1176 SD 10 16 30 35 55 65 76 145 161 232LFB-R565 + carboplatin Mean 87 139 184 212 200 421 480 810 756 936 SD 919 31 37 40 113 108 221 127 183 3C23K + carboplatin Mean 85 106 62 44 4056 40 53 46 114 111 165 SD 9 12 13 14 16 15 14 20 21 46 38 65

TABLE 4 Raw data for median tumour volumes and T/C ratios Day 11 14 1822 25 29 32 36 39 43 46 49 Mean tumour volume Control 91 163 216 392 441644 1020 1564 LFB-R565 81 154 173 270 410 693 690 1824 3C23K 80 84 72 7074 146 180 182 210 420 504 784 carboplatin 81 134 112 270 240 360 462630 660 882 LFB-R565 + carboplatin 81 150 180 210 170 330 385 704 691954 3C23K + carboplatin 83 96 53 24 23 40 18 18 24 45 55 86 TC ratio (%)Control-LFB-R565 89 95 80 69 93 108 68 117 Control-3C23K 88 52 33 18 1723 18 12 Control-carboplatin 89 82 52 69 54 56 45 40 Control-LFB-R565 +carboplatin 89 92 83 54 39 51 38 45 Control-3C23K + carboplatin 91 59 246 5 6 2 1 LFB-R565-3C23K 99 55 41 26 18 21 26 10 LFB-R565-carboplatin100 87 65 100 59 52 67 35 LFB-R565-LFB-R565 + carboplatin 100 97 104 7842 48 56 39 LFB-R565-3C23K + carboplatin 102 63 30 9 5 6 3 1carboplatin-3C23K 99 63 64 26 31 41 39 29 32 48 carboplatin-LFB-R565 +carboplatin 100 112 161 78 71 92 83 112 105 108 carboplatin-3C23K +carboplatin 102 72 47 9 9 11 4 3 4 5 3C23K-3C23K + carboplatin 103 11573 34 31 27 10 10 11 11 11 11 LFB-R565 + carboplatin-3C23K + carboplatin102 64 29 11 13 12 5 3 3 5

TABLE 5 Statistical analysis of the tumour volumes Statistical analysisof the tumour volumes Test Day Groups F ratio P value Sign. statistic Pvalue Sign. D11 Control-LFB-R565 0.13 0.73 0.44 0.51 Control-3C23K 0.130.72 0.33 0.57 Control-carboplatin 0.16 0.69 0.28 0.60Control-LFB-R565 + carboplatin 0.21 0.66 0.12 0.72 Control-3C23K +carboplatin 0.31 0.59 0.24 0.63 LFB-R565-3C23K 0.00 1.00 0.00 1.00LFB-R565-carboplatin 0.00 0.97 0.00 0.96 LFB-R565-LFB-R565 + carboplatin0.01 0.94 0.01 0.93 LFB-R565-3C23K + carboplatin 0.03 0.86 0.00 0.96carboplatin-3C23K 0.00 0.97 0.00 0.96 carboplatin-LFB-R565 + carboplatin0.00 0.97 0.00 1.00 carboplatin-3C23K + carboplatin 0.02 0.89 0.00 1.003C23K-LFB-R565 + carboplatin 0.01 0.94 0.01 0.93 3C23K-3C23K +carboplatin 0.03 0.86 0.00 0.96 LFB-R565 + carboplatin-3C23K +carboplatin 0.01 0.92 0.00 0.96 D14 Control-LFB-R565 1.23 0.28 1.22 0.27Control-3C23K 4.54 0.05 * 3.60 0.06 Control-carboplatin 0.64 0.44 0.570.45 Control-LFB-R565 + carboplatin 1.03 0.32 0.63 0.43 Control-3C23K +carboplatin 10.41 0.01 * 7.74 0.01 * LFB-R565-3C23K 1.57 0.23 1.88 0.17LFB-R565-carboplatin 0.05 0.83 0.10 0.76 LFB-R565-LFB-R565 + carboplatin0.01 0.93 0.20 0.66 LFB-R565-3C23K + carboplatin 3.96 0.06 2.98 0.08carboplatin-3C23K 1.88 0.19 2.00 0.16 carboplatin-LFB-R565 + carboplatin0.08 0.79 0.13 0.72 carboplatin-3C23K + carboplatin 4.26 0.06 3.780.05 * 3C23K-LFB-R565 + carboplatin 1.08 0.31 1.76 0.18 3C23K-3C23K +carboplatin 0.05 0.82 0.10 0.76 LFB-R565 + carboplatin-3C23K +carboplatin 2.39 0.14 1.13 0.29 D18 Control-LFB-R565 0.04 0.84 0.05 0.83Control-3C23K 25.29 0.00 * 11.57 0.00 * Control-carboplatin 1.51 0.242.26 0.13 Control-LFB-R565 + carboplatin 0.19 0.67 0.63 0.43Control-3C23K + carboplatin 25.53 0.00 * 10.96 0.00 * LFB-R565-3C23K14.01 0.00 * 11.62 0.00 * LFB-R565-carboplatin 1.39 0.26 2.27 0.13LFB-R565-LFB-R565 + carboplatin 0.29 0.59 0.20 0.66 LFB-R565-3C23K +carboplatin 14.50 0.00 * 11.01 0.00 * carboplatin-3C23K 8.37 0.01 * 9.590.00 * carboplatin-LFB-R565 + carboplatin 0.50 0.49 0.78 0.38carboplatin-3C23K + carboplatin 8.86 0.01 * 7.54 0.01 * 3C23K-LFB-R565 +carboplatin 13.69 0.00 * 9.84 0.00 * 3C23K-3C23K + carboplatin 0.07 0.800.20 0.66 LFB-R565 + carboplatin-3C23K + carboplatin 14.18 0.00 * 9.280.00 * D22 Control-LFB-R565 3.19 0.09 2.00 0.16 Control-3C23K 28.510.00 * 12.79 0.00 * Control-carboplatin 4.01 0.06 3.28 0.07Control-LFB-R565 + carboplatin 7.35 0.02 * 5.28 0.02 * Control-3C23K +carboplatin 36.82 0.00 * 12.86 0.00 * LFB-R565-3C23K 33.73 0.00 * 10.980.00 * LFB-R565-carboplatin 0.17 0.69 0.13 0.72 LFB-R565-LFB-R565 +carboplatin 2.12 0.16 1.76 0.18 LFB-R565-3C23K + carboplatin 52.970.00 * 12.87 0.00 * carboplatin-3C23K 22.58 0.00 * 10.39 0.00 *carboplatin-LFB-R565 + carboplatin 0.99 0.33 1.12 0.29carboplatin-3C23K + carboplatin 35.55 0.00 * 12.86 0.00 *3C23K-LFB-R565 + carboplatin 11.13 0.00 * 6.79 0.01 * 3C23K-3C23K +carboplatin 2.19 0.16 2.14 0.14 LFB-R565 + carboplatin-3C23K +carboplatin 19.45 0.00 * 11.62 0.00 * D25 Control-LFB-R565 1.02 0.330.44 0.51 Control-3C23K 31.60 0.00 * 12.84 0.00 * Control-carboplatin6.33 0.02 * 6.33 0.01 * Control-LFB-R565 + carboplatin 12.96 0.00 * 9.560.00 * Control-3C23K + carboplatin 42.71 0.00 * 12.84 0.00 *LFB-R565-3C23K 50.07 0.00 * 12.54 0.00 * LFB-R565-carboplatin 4.510.05 * 5.07 0.02 * LFB-R565-LFB-R565 + carboplatin 13.94 0.00 * 7.500.01 * LFB-R565-3C23K + carboplatin 83.99 0.00 * 12.84 0.00 *carboplatin-3C23K 9.51 0.01 * 8.27 0.00 * carboplatin-LFB-R565 +carboplatin 0.84 0.37 0.86 0.35 carboplatin-3C23K + carboplatin 16.390.00 * 11.61 0.00 * 3C23K-LFB-R565 + carboplatin 6.89 0.02 * 5.09 0.02 *3C23K-3C23K + carboplatin 1.54 0.23 0.73 0.39 LFB-R565 +carboplatin-3C23K + carboplatin 15.49 0.00 * 9.87 0.00 * D29Control-LFB-R565 0.13 0.72 0.00 1.00 Control-3C23K 28.90 0.00 * 12.800.00 * Control-carboplatin 11.64 0.00 * 9.57 0.00 * Control-LFB-R565 +carboplatin 4.78 0.04 * 5.28 0.02 * Control-3C23K + carboplatin 47.260.00 * 12.80 0.00 * LFB-R565-3C23K 56.20 0.00 * 12.83 0.00 *LFB-R565-carboplatin 18.53 0.00 * 9.86 0.00 * LFB-R565-LFB-R565 +carboplatin 5.30 0.04 * 4.31 0.04 * LFB-R565-3C23K + carboplatin 127.110.00 * 12.83 0.00 * carboplatin-3C23K 6.17 0.02 * 4.31 0.04 *carboplatin-LFB-R565 + carboplatin 0.39 0.54 0.03 0.86carboplatin-3C23K + carboplatin 21.04 0.00 * 12.82 0.00 *3C23K-LFB-R565 + carboplatin 5.32 0.03 * 4.13 0.04 * 3C23K-3C23K +carboplatin 3.64 0.07 1.44 0.23 LFB-R565 + carboplatin-3C23K +carboplatin 11.49 0.00 * 12.80 0.00 * D32 Control-LFB-R565 0.80 0.390.20 0.66 Control-3C23K 12.70 0.00 * 11.57 0.00 * Control-carboplatin7.03 0.02 * 9.56 0.00 * Control-LFB-R565 + carboplatin 6.13 0.02 * 7.250.01 * Control-3C23K + carboplatin 20.13 0.00 * 12.87 0.00 *LFB-R565-3C23K 18.68 0.00 * 10.40 0.00 * LFB-R565-carboplatin 8.060.01 * 6.33 0.01 * LFB-R565-LFB-R565 + carboplatin 5.95 0.03 * 5.480.02 * LFB-R565-3C23K + carboplatin 40.84 0.00 * 12.87 0.00 *carboplatin-3C23K 5.03 0.04 * 4.13 0.04 * carboplatin-LFB-R565 +carboplatin 0.02 0.88 0.05 0.83 carboplatin-3C23K + carboplatin 33.410.00 * 12.87 0.00 * 3C23K-LFB-R565 + carboplatin 3.99 0.06 3.13 0.08 *3C23K-3C23K + carboplatin 5.01 0.04 * 1.92 0.17 LFB-R565 +carboplatin-3C23K + carboplatin 18.20 0.00 * 12.87 0.00 * D36Control-LFB-R565 0.22 0.65 0.28 0.60 Control-3C23K 36.45 0.00 * 10.760.00 * Control-carboplatin 10.34 0.01 * 6.03 0.01 * Control-LFB-R565 +carboplatin 5.70 0.03 * 5.33 0.02 * Control-3C23K + carboplatin 70.670.00 * 12.07 0.00 * LFB-R565-3C23K 32.80 0.00 * 11.01 0.00 *LFB-R565-carboplatin 11.39 0.00 * 6.12 0.01 * LFB-R565-LFB-R565 +carboplatin 7.22 0.02 * 5.07 0.02 * LFB-R565-3C23K + carboplatin 54.440.00 * 12.86 0.00 * carboplatin-3C23K 8.19 0.01 * 5.50 0.02 *carboplatin-LFB-R565 + carboplatin 0.04 0.84 0.02 0.89carboplatin-3C23K + carboplatin 26.34 0.00 * 12.86 0.00 *3C23K-LFB-R565 + carboplatin 5.33 0.03 * 5.09 0.02 * 3C23K-3C23K +carboplatin 4.99 0.04 * 1.68 0.19 LFB-R565 + carboplatin-3C23K +carboplatin 13.13 0.00 * 12.86 0.00 * D39 carboplatin-3C23K 5.05 0.04 *4.70 0.03 * carboplatin-LFB-R565 + carboplatin 0.11 0.75 0.00 1.00carboplatin-3C23K + carboplatin 25.47 0.00 * 12.84 0.00 *3C23K-LFB-R565 + carboplatin 4.64 0.05 * 4.50 0.03 * 3C23K-3C23K +carboplatin 5.08 0.04 * 2.34 0.13 LFB-R565 + carboplatin-3C23K +carboplatin 39.24 0.00 * 12.06 0.00 * D43 carboplatin-3C23K 5.46 0.03 *4.32 0.04 * carboplatin-LFB-R565 + carboplatin 0.72 0.41 0.33 0.56carboplatin-3C23K + carboplatin 20.12 0.00 * 12.03 0.00 *3C23K-LFB-R565 + carboplatin 2.85 0.11 2.38 0.12 LFB-R565 +carboplatin-3C23K + carboplatin 21.66 0.00 * 10.63 0.00 * 3C23K-3C23K +carboplatin 4.91 0.04 * 1.34 0.25 D46 3C23K-3C23K + carboplatin 3.950.07 1.34 0.25 D49 3C23K-3C23K + carboplatin 5.02 0.04 * 1.34 0.25

TABLE 6 Summary of the raw data for individual body weight Individualbody weight of the mice Mouse Sep. 19, 2011 Sep. 26, 2011 Oct. 03, 2011Oct. 10, 2011 Oct. 17, 2011 Oct. 24, 2011 Group number D11 D18 D25 D32D39 D46 NaCl 1-SM 25.6 27.3 28.2 29.2 1-OD 22.3 24.8 24.7 25.0 27.0 28.91-OG 25.0 27.6 28.0 29.0 1-2O 21.5 22.8 23.2 24.6 25.8 1-2OD 25.0 27.028.1 29.6 29.6 2-SM 22.4 23.6 23.8 25.4 27.1 2-OD 24.0 26.1 26.8 29.330.0 2-OG 22.8 22.0 23.8 25.6 25.0 2-2O 24.4 24.4 25.2 27.0 27.1 29.6LFB-R565 3-SM 23.4 23.3 24.0 25.1 3-OD 22.2 23.6 23.8 23.9 25.2 27.03-OG 23.5 24.4 24.8 25.8 3-2O 21.5 21.8 22.3 23.5 24.6 3-2OD 25.4 25.626.4 28.2 4-SM 23.6 25.0 25.1 26.6 28.2 4-OD 27.1 26.4 28.2 29.7 30.530.7 4-OG 23.2 24.1 24.6 26.3 26.8 4-2O 24.9 26.0 26.0 29.7 30.4 32.83C23K 5-SM 21.0 22.1 22.1 22.6 23.1 22.5 5-OD 20.2 21.8 22.2 23.5 24.124.9 5-OG 25.8 26.6 26.6 27.0 27.2 27.2 5-2O 24.9 25.2 25.0 25.2 27.826.0 5-2OD 23.5 26.4 26.0 26.0 26.5 27.2 6-SM 21.4 21.8 22.3 23.0 23.823.3 6-OD 23.0 24.2 25.0 24.0 26.0 25.7 6-OG 23.9 24.5 25.8 25.8 27.928.9 6-2O 24.9 24.0 24.3 24.8 25.3 25.8 carboplatin 7-SM 22.3 22.2 27.623.7 24.3 25.4 7-OD 25.8 24.6 28.3 24.5 23.9 25.7 7-OG 23.0 22.6 25.623.6 25.1 26.4 7-2O 27.1 26.1 32.7 28.0 27.2 7-2OD 23.3 24.4 27.0 23.125.4 8-SM 24.5 25.4 29.4 23.1 22.4 25.7 8-OD 22.7 23.6 25.6 21.1 20.021.7 8-OG 20.4 20.3 24.2 20.2 21.0 21.0 8-2O 22.3 22.0 26.0 23.6 23.325.0 LFB-R565 + 9-SM 22.2 21.8 26.2 20.9 22.7 23.1 carboplatin 9-OD 22.422.9 26.9 18.3 22.3 22.8 9-OG 24.0 23.9 27.7 23.0 23.8 24.8 9-2O 18.919.2 23.5 22.2 19.1 19.8 9-2OD 21.5 22.9 27.1 21.8 10-SM 23.0 22.6 23.922.7 25.3 24.3 10-OD 25.2 25.4 26.6 26.1 25.6 27.8 10-OG 25.8 25.3 28.227.0 26.2 27.5 10-2O 23.4 23.3 23.6 21.4 18.6 3C23K + 11-SM 26.8 26.826.3 27.0 26.9 27.5 carboplatin 11-OD 21.3 21.5 22.6 22.8 21.4 23.211-OG 20.0 19.2 20.1 18.8 17.0 11-2O 24.0 24.8 23.1 24.0 23.1 25.011-2OD 24.6 23.5 22.9 25.2 22.5 24.0 12-SM 21.8 23.7 24.0 22.5 21.0 22.512-OD 22.3 20.5 23.7 22.7 22.3 23.7 12-OG 22.7 24.2 24.4 23.1 21.6 22.812-2O 22.8 21.9 25.0 23.6 23.6 25.2

TABLE 7 Summary of the raw data for mean body weight and standarddeviation (SD). Mean body weight of the mice Day Group 11 18 25 32 39 46Control Mean 23.7 25.1 25.8 27.2 SD 1.5 2.0 2.0 2.1 LFB-R565 Mean 23.924.5 25.0 26.5 SD 1.7 1.5 1.7 2.3 3C23K Mean 23.2 24.1 24.4 24.7 25.725.7 SD 1.9 1.9 1.8 1.5 1.8 2.0 carboplatin Mean 23.5 23.5 27.4 23.423.6 SD 2.0 1.8 2.5 2.2 2.2 LFB-R565 + Mean 22.9 23.0 26.0 22.6 23.0carboplatin SD 2.1 1.9 1.8 2.6 2.9 3C23K + carboplatin Mean 22.9 22.923.6 23.3 22.2 24.2 SD 2.0 2.3 1.7 2.2 2.6 1.6

2.2 Evaluation of an Anti-Tumour Activity of the 3C23K Antibody inCombination with Paclitaxel

The 3C23K antibody and paclitaxel are evaluated in vivo in monotherapyor in combination in the mice that received the injection ofCov434-AMHRII Asc1a5 tumour cells (a cell line of human ovarian cancer).The treatment is initiated on day 14 after injection.

It was observed that the 3C23K antibody administered at 10 mg/kgaccording to the Q7D4 regimen described above displays significantanti-tumour activity compared to a control solution (NaCl solution)(FIGS. 3 and 4; Tables 9, 11, 12, 13, 14 and 15).

Relative to the group treated with the control solution, the T/C ratiosof the group treated with the 3C23K antibody decreased starting fromD20. On D20 the T/C ratio was 57% and was constantly less than 29% up toD40 (Table 12).

Paclitaxel alone only shows a modest inhibition of tumour growthcompared to the control solution. From D24 to D41, the T/C ratio changesfrom 83% to 53%.

Compared to the control, a greater anti-tumour activity was obtained inthe group treated with the combination of the 3C23K antibody andpaclitaxel (FIGS. 3 and 4; Tables 9, 11, 12, 13 14 and 15). The T/Cratio was less than 38% on D20 and was always less than 28% up to D41(Table 12). Moreover, the T/C ratio on D31, D35 and D41 was 6%, 11% and12% respectively. Compared to the result described by Bissery et al. in1991, the results of the present invention show that the T/C ratio wasaround and less than 10% between D31 and D41, indicating that treatmentwith a combination of the 3C23K antibody and paclitaxel is an effectivetherapy.

Moreover, the 3C23K antibody administered at 10 mg/kg according to theQ7D4 regimen showed an anti-tumour effect greater than that ofpaclitaxel. In fact, the T/C ratio of the group treated with 3C23Krelative to the group treated with paclitaxel decreased starting fromD20. The T/C ratio on D20 was 47% and was less than 39% on D41 (Table12), which indicates an anti-tumour activity of 3C23K greater than thatof paclitaxel.

The combination of the 3C23K antibody (10 mg/kg, Q7D4) and paclitaxel(15 mg/kg, Q7D4 regimen) showed an anti-tumour effect stronger than eachof the components, 3C23K (10 mg/kg, Q7D4) or paclitaxel (15 mg/kg, Q7D4regimen) used in monotherapy.

In fact, when the group treated with the aforesaid combination wascompared with the group treated with paclitaxel, the combination showeda greater anti-tumour activity: on D20 the T/C ratio began to decreaseand the ratio was 31% on D20 and was less than 35% up to D41 (Table 12).

When the group treated with the aforesaid combination was compared withthe group treated with the 3C23K antibody, the combination showed ananti-tumour activity greater than that of monotherapy with 3C23K. OnD31, the T/C ratio began to decrease, the T/C ratio was 27% on D31 andwas less than 42% up to D48 (Table 12). On D51, the T/C ratio is only51%, nevertheless indicating a difference between these two groups, as49% inhibition of tumour growth was observed (Table 12).

Statistical analysis of tumour volume confirms the results of analysisof the T/C ratio.

The 3C23K antibody in monotherapy and the 3C23K antibody in combinationwith paclitaxel show a significant anti-tumour activity compared to thatof the control (NaCl solution) starting from D20 to D41 (Table 13).

Paclitaxel in monotherapy does not have an activity that issignificantly different from that of the control (Table 13).

The 3C23K antibody in monotherapy displays an anti-tumour activity thatis significantly different from that of paclitaxel in monotherapystarting from D20 to D41 (Table 13).

Compared to the paclitaxel treatment in monotherapy, treatment with thecombination of the 3C23K antibody and paclitaxel displays a betteranti-tumour activity starting from D20 to D41 (Table 13).

Statistical analysis based on the survival parameter also confirms theresults for the T/C ratio and analysis of tumour volume.

The 3C23K antibody in monotherapy and the combination of the 3C23Kantibody and paclitaxel respectively show a significant anti-tumouractivity compared to that of the control (Table 15).

Paclitaxel in monotherapy does not have an activity that issignificantly different from that of the control (Table 15).

The 3C23K antibody in monotherapy displays an anti-tumour activity thatis significantly different from that of paclitaxel in monotherapy (Table15). Compared to the paclitaxel treatment in monotherapy, treatment withthe combination of the 3C23K antibody and paclitaxel displays a betteranti-tumour activity (Table 15).

The treatments have no effect on body weight (Tables 8 and 10).

In conclusion, the combination of the 3C23K antibody and paclitaxeldisplays an advantage relative to 3C23K alone or to paclitaxel alone andas a minimum has an additive effect.

TABLE 8 Summary of the raw data for individual body weight Individualbody weight Mouse Group number D20 D27 D35 D41 D48 D55 D62 D69 Control1-SM 23.4 24.1 26.0 26.9 1-OD 23.6 25.4 26.5 1-OG 23.1 24.6 25.0 26.11-2O 21.8 23.4 25.2 25.4 1-2OG 23.7 25.8 26.8 27.8 1- 22.5 23.7 24.825.0 2OG/OD 10-2OD 22.1 23.4 24.3 23.5 24.7 26.1 10-2OG 22.4 23.5 28.427.8 28.5 29.6 3C23K 10 mg/kg 2-SM 21.3 21.8 23.0 22.4 23.3 24.6 q7d42-OD 25.0 24.8 26.8 27.0 27.5 2-OG 24.8 26.3 27.0 27.6 27.2 27.5 2-2O23.7 25.0 25.4 27.0 25.4 25.5 26.6 26.6 2-2OD 25.5 26.3 26.7 27.0 26.826.9 27.6 27.7 2-2OG 27.1 26.9 28.7 29.2 28.7 28.0 28.2 28.6 8-SM 20.521.0 21.4 21.7 22.0 22.4 23.4 8-OD 25.3 26.1 26.0 26.5 27.6 27.4 28.928.7 Paclitaxel 15 mg/kg 6-SM 25.6 27.1 30.7 0.0 0.0 0.0 0.0 0.0 q7d46-OD 25.8 25.7 27.8 27.7 28.0 0.0 0.0 0.0 6-OG 24.5 24.3 26.2 26.5 0.00.0 0.0 0.0 6-2O 24.4 24.6 27.7 27.2 0.0 0.0 0.0 0.0 6-2OD 22.4 24.026.5 26.7 0.0 0.0 0.0 0.0 6-2OG 25.9 26.4 29.0 28.7 0.0 0.0 0.0 0.010-SM 24.8 25.0 26.8 27.1 26.8 0.0 0.0 0.0 10-OD 23.9 23.6 23.9 23.725.2 26.3 0.0 0.0 3C23K 10 mg/kg 7-SM 22.0 22.4 22.3 22.3 25.5 22.2 0.00.0 q7d4 7-OD 26.4 26.8 26.8 25.7 26.7 28.3 29.4 0.0 + 7-OG 24.1 25.125.9 25.6 26.3 26.2 0.0 0.0 Paclitaxel 15 mg/kg 7-2O 22.0 22.2 23.7 23.425.1 0.0 0.0 0.0 q7d4 7-2OD 22.4 22.6 24.3 26.0 24.4 25.8 26.6 0.0 7-2OG22.5 23.6 25.0 25.0 24.9 26.7 27.6 0.0 10-OG 22.8 23.7 24.7 23.6 24.724.5 24.6 25.1 10-2O 22.7 23.7 24.4 23.8 24.7 25.0 25.4 26.8

TABLE 9 Summary of the raw data for individual tumour volume Day Group13 20 24 27 31 35 41 45 48 51 Control 1-SM 150 403 495 910 840 1566 19662320 1-OD 98 423 660 1190 1568 2898 1-OG 96 189 399 561 1344 1734 22001-2O 90 210 297 423 520 770 1584 2058 1-2OG 74 187 358 576 1056 12762474 1- 65 162 315 469 725 1089 1396 2128 10-2OD 106 72 41 50 24 117 197510 819 1008 10-2OG 66 105 81 126 143 366 631 725 1170 1521 3C23K 10mg/kg q7d4 2-SM 144 105 158 252 184 366 560 918 912 1485 2-OD 98 135 180289 404 592 844 1256 1672 2035 2-OG 98 88 0 0 0 0 0 0 0 0 2-2O 90 140102 192 204 488 540 624 551 587 2-2OD 76 150 114 168 198 528 634 731 833900 2-2OG 73 60 68 72 68 122 293 392 381 462 8-SM 88 105 84 90 140 268394 529 689 837 8-OD 72 108 72 90 28 119 169 166 276 351 Paclitaxel 15mg/kg q7d4 6-SM 126 424 756 896 1530 2706 6-OD 117 243 257 325 420 6991064 1632 1536 2166 6-OG 98 240 356 386 673 1200 2380 6-2O 83 280 284255 410 1008 1309 2040 6-2OD 83 170 380 560 720 1653 1389 2112 6-2OG 66126 180 347 410 965 2269 10-SM 144 217 180 270 358 823 884 1197 13232112 10-OD 58 105 110 198 192 351 437 798 1232 1348 3C23K 10 mg/kgq7d4 + 7-SM 131 126 85 0 0 0 0 0 0 0 Paclitaxel 15 mg/kg q7d4 7-OD 116105 112 223 160 324 315 390 655 938 7-OG 95 28 0 0 0 0 0 0 0 0 7-2O 8870 142 150 264 768 875 1033 1400 2112 7-2OD 74 41 112 105 163 357 368336 399 532 7-2OG 66 41 63 85 63 203 347 281 429 662 10-OG 110 96 104 960 0 0 0 0 0 10-2O 61 72 72 27 24 66 63 68 105 200 Day Group 55 58 62 6669 73 76 80 83 Control 1-SM 1-OD 1-OG 1-2O 1-2OG 1- 10-2OD 2268 10-2OG2145 3C23K 10 mg/kg q7d4 2-SM 2080 2-OD 2-OG 0 0 0 0 0 0 0 0 0 2-2O 9901482 1080 1695 1788 1820 2296 2-2OD 1084 1350 1440 1512 1496 1559 12381520 1615 2-2OG 600 842 640 666 950 1018 1140 1254 1463 8-SM 1309 16652057 8-OD 600 792 1215 1740 2000 Paclitaxel 15 mg/kg q7d4 6-SM 6-OD 6-OG6-2O 6-2OD 6-2OG 10-SM 10-OD 2132 3C23K 10 mg/kg q7d4 + 7-SM 0 0 0 0 0 00 0 0 Paclitaxel 15 mg/kg q7d4 7-OD 1755 1710 2340 7-OG 0 0 0 0 0 0 0 00 7-2O 7-2OD 816 1140 1170 2016 7-2OG 924 1109 1620 2081 10-OG 24 53 5060 96 189 192 347 413 10-2O 324 551 714 1040 1245 1604 1725 2112

TABLE 10 Summary of the raw data for change in mean body weight DaysGroups 20 27 35 41 48 55 control Mean 22.83 24.24 25.88 26.07 SD 0.720.94 1.34 1.57 3C23K 10 mg/kg q7d4 Mean 24.15 24.78 25.63 26.05 26.0626.04 SD 2.22 2.21 2.36 2.60 2.32 2.01 Paclitaxel 15 mg/kg Mean 24.6625.09 27.33 26.80 q7d4 SD 1.17 1.22 2.02 1.55 3C23K 10 mg/kg Mean 23.1123.76 24.64 24.43 25.29 25.53 q7d4 + Paclitaxel SD 1.48 1.54 1.36 1.330.82 1.91 15 mg/kg q7d4

TABLE 11 Summary of the raw data for the mean tumour volumes Days Groups13 20 24 27 31 35 41 45 48 51 55 Control Mean 93 219 331 538 777 12271492 SD 10 48 77 142 205 331 339 3C23K 10 mg/kg q7dx4 Mean 92 111 97 144153 310 429 577 664 832 SD 16 23 41 81 94 183 215 305 362 482 paclitaxel15 mg/kg q7dx4 Mean 97 226 313 405 589 1176 1390 SD 11 38 76 86 157 274290 3C23K 10 mg/kg + paclitaxel Mean 92 72 86 86 84 215 246 263 373 555549 15 mg/kg q7dx4 SD 9 13 16 29 38 101 114 132 182 272 269

TABLE 12 Summary of the raw data for the median tumour volumes and T/Cratios Days 13 20 24 27 31 35 41 45 48 51 55 Median tumour volumecontrol 93 188 336 515 782 1183 1584 3C23K 10 mg/kg q7dx4 89 107 93 129162 317 467 577 620 712 paclitaxel 15 mg/kg q7dx4 90 228 270 336 415 9861309 3C23K 10 mg/kg + paclitaxel 15 mg/kg q7dx4 91 71 94 90 44 134 189174 252 366 324 T/C ratio control-3C23K 10 mg/kg q7dx4 95 57 28 25 21 2729 control-paclitaxel 15 mg/kg q7dx4 97 121 80 65 53 83 83 control-3C23K10 mg/kg q7dx4 + paclitaxel 98 38 28 18 6 11 12 15 mg/kg q7dx4 3C23K 10mg/kg q7dx4-paclitaxel 15 mg/kg 98 47 35 38 39 32 36 q7dx4 3C23K 10mg/kg q7dx4-3C23K 10 mg/kg 103 67 101 70 27 42 40 30 41 51 q7dx4 +paclitaxel 15 mg/kg q7dx4 paclitaxel 15 mg/kg q7dx4-3C23K 10 mg/kg + 10131 35 27 11 14 14 paclitaxel 15 mg/kg q7dx4

TABLE 13 Statistical analysis ANOVA Kruskal - Wallis F- P- P- DayContrast between groups ratio value Sig. Test value Sig. D13control-3C23K 10 mg/kg q7d4 0.00 0.95 0.00 0.96 control-paclitaxel 15mg/kg q7d4 0.07 0.80 0.04 0.83 control-3C23K 10 mg/kg q7d4 + paclitaxel0.00 0.97 0.00 1.00 15 mg/kg q7d4 3C23K 10 mg/kg q7d4-paclitaxel 15mg/kg 0.11 0.74 0.03 0.87 q7d4 3C23K 10 mg/kg q7d4-3C23K 10 mg/kg 0.000.98 0.00 0.96 q7d4 + paclitaxel 15 mg/kg q7d4 paclitaxel 15 mg/kgq7d4-3C23K 10 mg/kg 0.09 0.76 0.07 0.79 q7d4 + paclitaxel 15 mg/kg q7d4D20 control-3C23K 10 mg/kg q7d4 5.33 0.04 * 4.44 0.04 *control-paclitaxel 15 mg/kg q7d4 0.01 0.91 0.47 0.49 control-3C23K 10mg/kg q7d4 + paclitaxel 9.71 0.01 * 8.08 0.00 * 15 mg/kg q7d4 3C23K 10mg/kg q7d4-paclitaxel 15 mg/kg 9.54 0.01 * 6.39 0.01 * q7d4 3C23K 10mg/kg q7d4-3C23K 10 mg/kg 5.84 0.03 * 4.44 0.04 * q7d4 + paclitaxel 15mg/kg q7d4 paclitaxel 15 mg/kg q7d4-3C23K 10 mg/kg 16.67 0.00 * 9.970.00 * q7d4 + paclitaxel 15 mg/kg q7d4 D24 control-3C23K 10 mg/kg q7d49.83 0.01 * 4.41 0.04 * control-paclitaxel 15 mg/kg q7d4 0.03 0.86 0.280.60 control-3C23K 10 mg/kg q7d4 + paclitaxel 11.11 0.00 * 4.42 0.04 *15 mg/kg q7d4 3C23K 10 mg/kg q7d4-paclitaxel 15 mg/kg 8.53 0.01 * 8.700.00 * q7d4 3C23K 10 mg/kg q7d4-3C23K 10 mg/kg 0.19 0.67 0.10 0.75q7d4 + paclitaxel 15 mg/kg q7d4 paclitaxel 15 mg/kg q7d4-C23K 10 mg/kg9.71 0.01 * 9.30 0.00 * q7d4 + paclitaxel 15 mg/kg q7d4 D27control-3C23K 10 mg/kg q7d4 8.21 0.01 * 4.87 0.03 * control-paclitaxel15 mg/kg q7d4 0.74 0.40 1.10 0.29 control-3C23K 10 mg/kg q7d4 +paclitaxel 11.09 0.00 * 6.90 0.01 * 15 mg/kg q7d4 3C23K 10 mg/kgq7d4-paclitaxel 15 mg/kg 8.92 0.01 * 8.66 0.00 * q7d4 3C23K 10 mg/kgq7d4-3C23K 10 mg/kg 1.75 0.21 1.11 0.29 q7d4 + paclitaxel 15 mg/kg q7d4paclitaxel 15 mg/kg q7d4-3C23K 10 mg/kg 14.22 0.00 * 10.61 0.00 * q7d4 +paclitaxel 15 mg/kg q7d4 D31 control-3C23K 10 mg/kg q7d4 10.00 0.01 *4.86 0.03 * control-paclitaxel 15 mg/kg q7d4 0.61 0.45 0.89 0.34control-3C23K 10 mg/kg q7d4 + paclitaxel 12.60 0.00 * 6.67 0.01 * 15mg/kg q7d4 3C23K 10 mg/kg q7d4-paclitaxel 15 mg/kg 8.01 0.01 * 8.660.00 * q7d4 3C23K 10 mg/kg q7d4-3C23K 10 mg/kg 1.44 0.25 1.75 0.19q7d4 + paclitaxel 15 mg/kg q7d4 paclitaxel 15 mg/kg q7d4-3C23K 10 mg/kg11.13 0.00 * 10.68 0.00 * q7d4 + paclitaxel 15 mg/kg q7d4 D35control-3C23K 10 mg/kg q7d4 8.26 0.01 * 5.11 0.02 * control-paclitaxel15 mg/kg q7d4 0.02 0.90 0.10 0.75 control-3C23K 10 mg/kg q7d4 +paclitaxel 9.78 0.01 * 8.09 0.00 * 15 mg/kg q7d4 3C23K 10 mg/kgq7d4-paclitaxel 15 mg/kg 10.47 0.01 * 8.65 0.00 * q7d4 3C23K 10 mg/kgq7d4-3C23K 10 mg/kg 0.62 0.45 1.23 0.27 q7d4 + paclitaxel 15 mg/kg q7d4paclitaxel 15 mg/kg q7d4-3C23K 10 mg/kg 12.39 0.00 * 9.33 0.00 * q7d4 +paclitaxel 15 mg/kg q7d4 D41 control-3C23K 10 mg/kg q7d4 11.82 0.00 *5.36 0.02 * control-paclitaxel 15 mg/kg q7d4 0.06 0.81 0.20 0.65control-3C23K 10 mg/kg q7d4 + paclitaxel 15.82 0.00 * 7.14 0.01 * 15mg/kg q7d4 3C23K 10 mg/kg q7d4-paclitaxel 15 mg/kg 12.64 0.00 * 7.710.01 * q7d4 3C23K 10 mg/kg q7d4-3C23K 10 mg/kg 1.63 0.22 1.75 0.19q7d4 + paclitaxel 15 mg/kg q7d4 paclitaxel 15 mg/kg q7d4-3C23K 10 mg/kg17.31 0.00 * 9.83 0.00 * q7d4 + paclitaxel 15 mg/kg q7d4 D45 3C23K 10mg/kg q7d4-3C23K 10 mg/kg 2.75 0.12 3.05 0.08 q7d4 + paclitaxel 15 mg/kgq7d4 D48 3C23K 10 mg/kg q7d4-3C23K 10 mg/kg 1.38 0.26 1.75 0.19 q7d4 +paclitaxel 15 mg/kg q7d4 D51 3C23K 10 mg/kg q7d4-3C23K 10 mg/kg 0.650.43 0.81 0.37 q7d4 + paclitaxel 15 mg/kg q7d4

TABLE 14 Summary of the raw data for survival parameter Mouse numberGroups Days Observations 1 NaCl 34 2 NaCl 41 3 NaCl 41 4 NaCl 45 5 NaCl45 6 NaCl 45 7 NaCl 55 8 NaCl 55 1 3C23K 10 mg/kg q7d4 51 2 3C23K 10mg/kg q7d4 55 3 3C23K 10 mg/kg q7d4 62 4 3C23K 10 mg/kg q7d4 69 5 3C23K10 mg/kg q7d4 76 6 3C23K 10 mg/kg q7d4 83 End of experiment TV 1615 mm37 3C23K 10 mg/kg q7d4 83 End of experiment TV 1463 mm3 8 3C23K 10 mg/kgq7d4 83 End of experiment no tumour 1 Paclitaxel 15 mg/kg q7d4 34 2Paclitaxel 15 mg/kg q7d4 41 3 Paclitaxel 15 mg/kg q7d4 41 4 Paclitaxel15 mg/kg q7d4 45 5 Paclitaxel 15 mg/kg q7d4 45 6 Paclitaxel 15 mg/kgq7d4 51 7 Paclitaxel 15 mg/kg q7d4 51 8 Paclitaxel 15 mg/kg q7d4 55 13C23K 10 mg/kg q7d4 + Paclitaxel 51 15 mg/kg q7d4 2 3C23K 10 mg/kgq7d4 + Paclitaxel 62 15 mg/kg q7d4 3 3C23K 10 mg/kg q7d4 + Paclitaxel 6615 mg/kg q7d4 4 3C23K 10 mg/kg q7d4 + Paclitaxel 66 15 mg/kg q7d4 53C23K 10 mg/kg q7d4 + Paclitaxel 80 15 mg/kg q7d4 6 3C23K 10 mg/kgq7d4 + Paclitaxel 83 End of experiment 15 mg/kg q7d4 TV 413 mm3 7 3C23K10 mg/kg q7d4 + Paclitaxel 83 End of experiment 15 mg/kg q7d4 No tumour8 3C23K 10 mg/kg q7d4 + Paclitaxel 83 End of experiment 15 mg/kg q7d4 Notumour

TABLE 15 Survival parameters Survival parameter Group Mean Median SDControl 45 45 5 3C23K 10 mg/kg q7dx4 70 73 11 Paclitaxel 15 mg/kg q7dx445 45 5 3C23K 10 mg/kg + Paclitaxel 72 72 11 15 mg/kg q7dx4 Survivalparameter: logrank test chi-squared p-value sign. Control-3C23K 10 mg/kgq7d4 11.15 0.00 * Control-paclitaxel 15 mg/kg q7d4 0.04 0.84Control-3C23K 10 mg/kg q7d4 + 12.87 0.00 * paclitaxel 15 mg/kg q7d43C23K 10 mg/kg q7d4-paclitaxel 12.36 0.00 * 15 mg/kg q7d4 3C23K 10 mg/kgq7d4-3C23K 10 0.17 0.68 mg/kg q7d4 + paclitaxel 15 mg/kg q7d4 paclitaxel15 mg/kg q7d4-3C23K 10 13.61 0.00 * mg/kg q7d4 + paclitaxel 15 mg/kgq7d4

1. Pharmaceutical composition comprising, as active ingredient, incombination with a pharmaceutically acceptable vehicle, an anticanceragent, and an antibody binding the human anti-Müllerian hormone type IIreceptor (AMHR-II).
 2. Pharmaceutical composition according to claim 1,in which said antibody is a polyclonal antibody or a monoclonalantibody, and preferably a chimeric 12G4 or humanized 12G4 monoclonalantibody.
 3. Composition according to claim 1, in which said monoclonalantibody is a humanized 12G4 antibody, or a fragment of humanized 12G4monoclonal antibody, in which said humanized 12G4 monoclonal antibody ismutated and comprises at least one mutation in the light and/or heavychain, and said mutated humanized 12G4 monoclonal antibody in particularpossesses affinity for AMHR-II characterized by a K_(D) preferably lessthan 10⁻⁷ M, in particular less than 10⁻⁸ M, in particular in the rangefrom 10⁻⁹ M to 10⁻¹¹ M.
 4. Pharmaceutical composition according to claim3, in which said humanized 12G4 monoclonal antibody comprises or isconstituted by: a) a light chain comprising or constituted by: avariable region the amino acid sequence of which is represented by SEQID NO: 1 or SEQ ID NO: 2, and a constant region the amino acid sequenceof which is represented by SEQ ID NO: 3 or by a sequence having at least80% homology with SEQ ID NO: 3 b) a heavy chain comprising orconstituted by: a variable region the amino acid sequence of which isrepresented by SEQ ID NO: 4, or SEQ ID NO: 5, and a constant region theamino acid sequence of which is represented by SEQ ID NO: 6 or by asequence having at least 80% homology with SEQ ID NO: 6, in which thehumanized 12G4 monoclonal antibody comprises at least one mutation inthe light and/or heavy chain, and has a K_(D) for the humananti-Müllerian hormone type II receptor (AMHR-II) preferably less than10⁻⁷ M, in particular less than 10⁻⁸ M, in particular in the range from10⁻⁹ M to 10⁻¹¹ M.
 5. Pharmaceutical composition according to claim 4,in which said mutated humanized 12G4 monoclonal antibody comprises or isconstituted by: a) a light chain comprising or constituted by: avariable region the amino acid sequence of which is represented by SEQID NO: 7 or SEQ ID NO: 8, a constant region the amino acid sequence ofwhich is represented by SEQ ID NO: 3 b) a heavy chain comprising orconstituted by: a variable region the amino acid sequence of which isrepresented by SEQ ID NO: 9 or SEQ ID NO: 10 a constant region the aminoacid sequence of which is represented by SEQ ID NO: 6
 6. Pharmaceuticalcomposition according to claim 4, in which the mutated humanized 12G4monoclonal antibody is produced by the 3C23K clone or in which thefragment of humanized 12G4 monoclonal antibody is a fragment of themutated humanized 12G4 monoclonal antibody produced by the 3C23K clone.7. Pharmaceutical composition according to claim 1, in which theantibody is a recombinant antibody produced by animal transgenesis. 8.Pharmaceutical composition according to claim 1, in which the anticanceragent is paclitaxel or a platinum salt selected from the groupconsisting of: oxaliplatin, cisplatin, carboplatin, in particularcarboplatin.
 9. Pharmaceutical composition according to claim 8,comprising the mutated 12G4 monoclonal antibody produced by the 3C23Kclone, and carboplatin.
 10. Pharmaceutical composition according toclaim 8, comprising the mutated 12G4 monoclonal antibody produced by the3C23K clone, and paclitaxel.
 11. Pharmaceutical composition according toclaim 1, in a formulation intended for administration by the intravenousor intraperitoneal route.
 12. Pharmaceutical composition according toclaim 1, in which the therapeutically effective quantity of antibodyadministered to a patient is in a range from about 0.07 mg to about35000 mg, preferably from about 0.7 mg to about 7000 mg, preferably fromabout 0.7 mg to about 1400 mg, preferably from about 0.7 mg to about 700mg, and more preferably from about 0.7 mg to about 70 mg. 13.Pharmaceutical composition according to claim 1, in which thetherapeutically effective quantity of anticancer agent administered to apatient is in a range from about 10 mg to about 700 mg, preferably in arange from about 20 mg to about 350 mg, and preferably is about 110 mg.14. Pharmaceutical composition according to claim 1, in which the doseof antibody administered to a patient is about 70 mg and the dose ofanticancer agent administered to the patient is about 110 mg. 15.Composition comprising an anticancer agent and an antibody binding thehuman anti-Müllerian hormone type II receptor (AMHR-II), for use as amedicinal product in the prevention or treatment of a pathologyassociated with the human anti-Müllerian hormone type II receptor(AMHR-II).
 16. Composition for use as a medicinal product, according toclaim 15, in which the pathology associated with the humananti-Müllerian hormone type II receptor (AMHR-II) is cancer, andespecially ovarian cancer.
 17. Composition according to claim 2, inwhich said monoclonal antibody is a humanized 12G4 antibody, or afragment of humanized 12G4 monoclonal antibody, in which said humanized12G4 monoclonal antibody is mutated and comprises at least one mutationin the light and/or heavy chain, and said mutated humanized 12G4monoclonal antibody in particular possesses affinity for AMHR-IIcharacterized by a K_(D) preferably less than 10⁻⁷ M, in particular lessthan 10⁻⁸ M, in particular in the range from 10⁻⁹ M to 10⁻¹¹ M. 18.Pharmaceutical composition according to claim 5, in which the mutatedhumanized 12G4 monoclonal antibody is produced by the 3C23K clone or inwhich the fragment of humanized 12G4 monoclonal antibody is a fragmentof the mutated humanized 12G4 monoclonal antibody produced by the 3C23Kclone.